Peloruside Analogs

ABSTRACT

The present invention provides new therapeutic compounds for treatment of diseases caused or influenced by microtubule stability, such as proliferative diseases. The compounds are phenyl-analogs of the naturally occurring peloruside. The analogs are easier to synthesize, and hence open up the possibility to produce said compounds in large quantities for therapeutic use.

FIELD OF THE INVENTION

The present invention is situated in the field of medical treatment,more particularly in the field of treatment of microtubule-relateddisorders, particularly cancer treatment, using analogs of the naturalcompound peloruside with improved treatment characteristics. Theinvention relates to said peloruside analogs as such, and methodsinvolving the use of said peloruside analogs in treatment.

BACKGROUND OF THE INVENTION

New and potent drugs are urgently needed to inhibit uncontrolled growth,invasion and metastasis. The majority of available treatments ortherapeutics are limited towards inhibition of the growth of the tumour.Microtubule inhibitors block the mitotic spindle and function of thecytoplasmic microtubule complex and therefore form an interestingalternative for drug development for treating cancer, since they inhibitproliferation, invasion and metastasis. Two examples of such drugs thathave been used with success are paclitaxel (Taxol®) and docetaxel(Taxotere®), which have been used to treat roughly one million ofpatients in the 10 years since they were first approved as anti-cancerdrugs. Paclitaxel, docetaxel and vinca alkaloids like vinblastine arecompounds interacting with the mitotic spindle by binding to β-tubulin.They are used as therapeutics in standard chemotherapy regimens and incombination with new drugs like the HER2 targeting antibody trastuzumab.But toxicity, drug resistance, and complex galenic formulations arerestricting their clinical use in cancer therapy. Further drawbacks ofe.g. paclitaxel are its low solubility in water (needs to be dissolvedin Cremophor EL (polyoxyethylated castor oil) which causeshypersensitivity reactions), high hydrophobicity, adhesion to plastictubing inner surfaces complicating administration, creating therapyresistance, etc. . . . .

Thus, new tubulin targeting antimitotic agents with better tolerabilityand efficacy against (late-stage resistant) tumors are urgently needed.The present invention provides solutions to the above-identifiedproblems by designing peloruside-derivatives as new therapeutic agents.

SUMMARY OF THE INVENTION

The present invention follows from a study of peloruside. Peloruside wasfirst discovered in marine sponges in New Zealand, and comes mainly intwo forms, Peloruside A (West et al. J. Org. Chem., 2000, 65, 445-449)and B (Singh et al. J. Org. Chem. 2009, 75, 2-10):

Peloruside functions as an antitumor agent by promoting tubulinpolymerization and interfering with microtubule dynamics, which causesarrest of the cell cycle in the G₂-M phase, followed by apoptosis (Hoodet al. Cancer Res. 2002, 62, 3356-3360), even at nanomolarconcentrations. This mode of action is similar to that of paclitaxel.Peloruside however is less susceptible than paclitaxel to multidrugresistance, arising from overexpression of the P-glycoprotein (Gaitanoset al. Cancer Res., 2004, 64, 5063-5067). Moreover, it is proven thatpeloruside and paclitaxel do not compete for the same binding site.Hence, peloruside conserves its cytotoxicity in cell lines that areaffected by mutations of the β-tubuline gene, causing a conformationalchange of the taxoid binding site. Furthermore, it is proven thatpeloruside exhibits synergy with other microtubule stabilizing drugslike taxol (Hamel et al. Cancer Chemoth. Pharm., 2006, 70, 1555-1564).

The biggest drawback of using peloruside is that it is only found inmilligram quantities in a specific species of sea sponges with lownatural abundance.

The inventors designed peloruside analogs by replacing the pyranosemoiety with a phenyl moiety and could determine that its activity as amicrotubule inhibitor was comparable to that of Paclitaxel. On a weightper volume base the peloruside analogs are at least as potent aspaclitaxel in their ability to disturb the cytoplasmic microtubulecomplex. The smaller molecular weight of the former versus the lattermay be associated with better penetration through the cell membrane,with reduced aspecific cytotoxicity and escape from resistancemechanisms. Adhesion to plastic tubing inner surfaces, which cancomplicate paclitaxel administration, also seems to be reduced.

By using a phenyl ring to replace the pyranose ring, the number ofstereocenters is reduced from 10 to 6 and hence also the syntheticcomplexity is reduced, while retaining the biological activity.

The invention therefore provides compounds of Formula I, and thestereoisomers, prodrugs, tautomers, racemates, salts, hydrates, orsolvates thereof,

-   -   wherein    -   X¹ is CR^(1a)R^(1b), X² is CR^(2a)R^(2b), X³ is CR^(3a)R^(3b),        X⁴ is CR^(4a)R^(4b), wherein    -   R^(1a), R^(2a), R^(3a), and R^(4a) are each independently        selected from hydrogen, hydroxyl, halogen, and a group selected        from —NR¹⁰R¹¹, C₁₋₆alkyl, C₂₋₆alkenyl, C₂₋₆alkynyl, C₆₋₁₀aryl,        C₆₋₁₀arylC₁₋₆alkyl and C₁₋₆alkoxy, each group independently        being optionally substituted with one or more substituent(s)        each independently selected from hydroxyl, halogen, C₁₋₆alkyl        and C₁₋₆alkoxy; and    -   R^(1b), R^(2b), R^(3b), and R^(4b) each independently are        selected from hydrogen, hydroxyl, halogen, and a group selected        from —NR¹⁰R¹¹, C₁₋₆alkyl, C₂₋₆alkenyl, C₂₋₆alkynyl, C₆₋₁₀aryl,        C₆₋₁₀arylC₁₋₆alkyl and C₁₋₆alkoxy, each group independently        being optionally substituted with one or more substituent(s)        each independently selected from hydroxyl, halogen, C₁₋₆alkyl        and C₁₋₆alkoxy; or    -   R^(1a) and R^(1b), or R^(2a) and R^(2b), or R^(3a) and R^(3b),        or R^(4a) and R^(4b) taken together represent an oxo (═O) group;    -   wherein the bond represented by a dashed and solid line        represents a single bond or a double bond and in case of a        double bond, R^(1b) and R^(2b) are absent and at least one of        R^(1a) and R^(2a) is not OH or —NR¹⁰R¹¹; and    -   R¹⁰ and R¹¹ are each independently selected from hydrogen and        C₁₋₆alkyl; and wherein:        -   when R^(1a), R^(2a), R^(3a) or R^(4a) is hydroxyl, the            corresponding R^(1b), R^(2b), R^(3b) or R^(4b) is not            hydroxyl, is not —NR¹⁰R¹¹, is not halogen, or is not            C₁₋₆alkoxy,        -   when R^(1a), R^(2a), R^(3a) or R^(4a) is —NR¹⁰R¹¹, the            corresponding R^(1b), R^(2b), R^(3b) or R^(4b) is not            hydroxyl, is not —NR¹⁰R¹¹, is not halogen, or is not            C₁₋₆alkoxy,        -   when R^(1a), R^(2a), R^(3a) or R^(4a) is halogen, the            corresponding R^(1b), R^(2b), R^(3b) or R^(4b) is not            hydroxyl, or is not —NR¹⁰R¹¹,        -   when R^(1b), R^(2b), R^(3b) or R^(4b) is hydroxyl, the            corresponding R^(1a), R^(2a), R^(3a) or R^(4a) is not            hydroxyl, is not —NR¹⁰R¹¹, is not halogen, or is not            C₁₋₆alkoxy, and        -   when R^(1b), R^(2b), R^(3b) or R^(4b) is —NR¹⁰R¹¹, the            corresponding R^(1a), R^(2a), R^(3a) or R^(4a) is not            hydroxyl, is not —NR¹⁰R¹¹, is not halogen, or is not            C₁₋₆alkoxy.        -   when R^(1b), R^(2b), R^(3b) or R^(4b) is halogen, the            corresponding R^(1a), R^(2a), R^(3a) or R^(4a) is not            hydroxyl, or is not —NR¹⁰R¹¹,

Preferably, the invention provides compounds of Formula I, and thestereoisomers, prodrugs, tautomers, racemates, salts, hydrates, orsolvates thereof,

-   -   wherein:        -   X¹ is CR^(1a)R^(1b), X² is CR^(2a)R^(2b), X³ is            CR^(3a)R^(3b), X⁴ is CR^(4a)R^(4b), and wherein:        -   R^(1a) is selected from hydrogen, hydroxyl, halogen, and a            group selected from —NR¹⁰R¹¹, C₁₋₆alkyl, C₂₋₆alkenyl,            C₂₋₆alkynyl, C₆₋₁₀aryl, C₆₋₁₀arylC₁₋₆alkyl and C₁₋₆alkoxy,            each group being independently optionally substituted with            one or more substituent(s) each independently selected from            hydroxyl, halogen, C₁₋₆alkyl and C₁₋₆alkoxy;        -   R^(2a) is selected from hydrogen, hydroxyl, halogen, and a            group selected from —NR¹⁰R¹¹, C₁₋₆alkyl, C₂₋₆alkenyl,            C₂₋₆alkynyl, C₆₋₁₀aryl, C₆₋₁₀arylC₁₋₆alkyl and C₁₋₆alkoxy,            each group being independently optionally substituted with            one or more substituent(s) each independently selected from            hydroxyl, halogen, C₁₋₆alkyl and C₁₋₆alkoxy,        -   R^(3a) is selected from hydrogen, hydroxyl, halogen, and a            group selected from —NR¹⁰R¹¹, C₁₋₆alkyl, C₂₋₆alkenyl,            C₂₋₆alkynyl, C₆₋₁₀aryl, C₆₋₁₀arylC₁₋₆alkyl and C₁₋₆alkoxy,            each group being independently optionally substituted with            one or more substituent(s) each independently selected from            hydroxyl, halogen, C₁₋₆alkyl and C₁₋₆alkoxy        -   R^(4a) is selected from hydrogen, hydroxyl, halogen, and a            group selected from —NR¹⁰R¹¹, C₁₋₆alkyl, C₂₋₆alkenyl,            C₂₋₆alkynyl, C₆₋₁₀aryl, C₆₋₁₀arylC₁₋₆alkyl and C₁₋₆alkoxy,            each group being independently optionally substituted with            one or more substituent(s) each independently selected from            hydroxyl, halogen, C₁₋₆alkyl and C₁₋₆alkoxy; and wherein:    -   R^(1b) is selected from hydrogen, hydroxyl, halogen, and a group        selected from —NR¹⁰R¹¹, C₁₋₆alkyl, C₂₋₆alkenyl, C₂₋₆alkynyl,        C₆₋₁₀aryl, C₆₋₁₀arylC₁₋₆alkyl and C₁₋₆alkoxy, each group being        independently optionally substituted with one or more        substituent(s) each independently selected from hydroxyl,        halogen, C₁₋₆alkyl and C₁₋₆alkoxy,        -   R^(2b) is selected from hydrogen, hydroxyl, halogen, and a            group selected from —NR¹⁰R¹¹, C₁₋₆alkyl, C₂₋₆alkenyl,            C₂₋₆alkynyl, C₆₋₁₀aryl, C₆₋₁₀arylC₁₋₆alkyl and C₁₋₆alkoxy,            each group being independently optionally substituted with            one or more substituent(s) each independently selected from            hydroxyl, halogen, C₁₋₆alkyl and C₁₋₆alkoxy,        -   R^(3b) is selected from hydrogen, hydroxyl, halogen, and a            group selected from —NR¹⁰R¹¹, C₁₋₆alkyl, C₂₋₆alkenyl,            C₂₋₆alkynyl, C₆₋₁₀aryl, C₆₋₁₀arylC₁₋₆alkyl and C₁₋₆alkoxy,            each group being independently optionally substituted with            one or more substituent(s) each independently selected from            hydroxyl, halogen, C₁₋₆alkyl and C₁₋₆alkoxy,        -   R^(4b) is selected from hydrogen, hydroxyl, halogen, and a            group selected from —NR¹⁰R¹¹, C₁₋₆alkyl, C₂₋₆alkenyl,            C₂₋₆alkynyl, C₆₋₁₀aryl, C₆₋₁₀arylC₁₋₆alkyl and C₁₋₆alkoxy,            each group independently being independently optionally            substituted with one or more substituent(s) each            independently selected from hydroxyl, halogen, C₁₋₆alkyl and            C₁₋₆alkoxy; or wherein R^(1a) and R^(1b), or R^(2a) and            R^(2b), or R^(3a) and R^(3b), or R^(4a) and R^(4b) taken            together represent an oxo (═O) group; and wherein the bond            represented by a dashed and solid line represents a single            bond or a double bond and wherein in case of a double bond,            R^(1b) and R^(2b) are absent and at least one of R^(1a) and            R^(2a) is not OH or —NR¹⁰R¹¹; and wherein:        -   R¹⁰ and R¹¹ are each independently selected from hydrogen            and C₁₋₆alkyl; and wherein:        -   when R^(1a), R^(2a), R^(3a) or R^(4a) is hydroxyl, the            corresponding R^(1b), R^(2b), R^(3b) or R^(4b) is not            hydroxyl, is not —NR¹⁰R¹¹, is not halogen, or is not            C₁₋₆alkoxy,        -   when R^(1a), R^(2a), R^(3a) or R^(4a) is —NR¹⁰R¹¹, the            corresponding R^(1b), R^(2b), R^(3b) or R^(4b) is not            hydroxyl, is not —NR¹⁰R¹¹, is not halogen, or is not            C₁₋₆alkoxy,        -   when R^(1a), R^(2a), R^(3a) or R^(4a) is halogen, the            corresponding R^(1b), R^(2b), R^(3b) or R^(4b) is not            hydroxyl, or is not —NR¹⁰R¹¹,        -   when R^(1b), R^(2b), R^(3b) or R^(4b) is hydroxyl, the            corresponding R^(1a), R^(2a), R^(3a) or R^(4a) is not            hydroxyl, is not —NR¹⁰R¹¹, is not halogen, or is not            C₁₋₆alkoxy, and        -   when R^(1b), R^(2b), R^(3b) or R^(4b) is —NR¹⁰R¹¹, the            corresponding R^(1a), R^(2a), R^(3a) or R^(4a) is not            hydroxyl, is not —NR¹⁰R¹¹, is not halogen, or is not            C₁₋₆alkoxy.        -   when R^(1b), R^(2b), R^(3b) or R^(4b) is halogen, the            corresponding R^(1a), R^(2a), R^(3a) or R^(4a) is not            hydroxyl, or is not —NR¹⁰R¹¹,

Preferably, in the structures of general formula I as defined above,R^(1b), R^(2b), R^(3b), and R^(4b) each independently are selected fromhydrogen and halogen and a group selected from C₁₋₆alkyl, C₂₋₆alkenyl,C₂₋₆alkynyl, C₆₋₁₀aryl, C₆₋₁₀arylC₁₋₆alkyl, and C₁₋₆alkoxy, each groupbeing optionally substituted with one or more substituent(s) selectedfrom hydroxyl, halogen, C₁₋₆alkyl, or C₁₋₆alkoxy; or wherein R^(1a) andR^(1b), and/or R^(2a) and R^(2b), and/or R^(3a) and R^(3b), and/orR^(4a) and R^(4b) taken together represent an oxo (═O) group.

In a preferred embodiment, in the structures of general formula I asdefined above, said R^(1b), R^(2b), R^(3b), and R^(4b) eachindependently are selected from hydrogen, halogen, and a group selectedfrom C₁₋₆alkyl, C₂₋₆alkenyl, C₂₋₆alkynyl, C₆₋₁₀aryl, C₆₋₁₀arylC₁₋₆alkyl,and C₁₋₆alkoxy, each group being optionally substituted with one or moresubstituent(s) selected from hydroxyl, halogen, C₁₋₆alkyl, orC₁₋₆alkoxy.

In a preferred embodiment, in the structures of general formula I asdefined above, said R^(1b), R^(2b), and R^(3b) each independently areselected from hydrogen, halogen, and a group selected from C₁₋₆alkyl,C₂₋₆alkenyl, C₂₋₆alkynyl, C₆₋₁₀aryl, C₆₋₁₀arylC₁₋₆alkyl, and C₁₋₆alkoxy,each group being optionally substituted with one or more substituent(s)selected from hydroxyl, halogen, C₁₋₆alkyl, or C₁₋₆alkoxy, and whereinR^(4a) and R^(4b) taken together represent an oxo (═O) group.

In a preferred embodiment, the invention provides compounds havingstructural Formula Ia, and the stereoisomers, prodrugs, tautomers,racemates, salts, hydrates, or solvates thereof:

wherein R^(1b), R^(2b), R^(3b), and R^(4b) have the same meaning asdefined above.

In another preferred embodiment, the invention provides compounds havingstructural Formula Ib, Ic or Id, and the stereoisomers, prodrugs,tautomers, racemates, salts, hydrates, or solvates thereof:

wherein R^(1b), R^(2b), R^(3b), and R^(4b) have the same meaning asdefined above. The double bond in structure Ic can have the Z- orE-geometry.

In another preferred embodiment, the invention provides compounds havingstructural Formula Ie, If, or Ig, and the stereoisomers, prodrugs,tautomers, racemates, salts, hydrates, or solvates thereof:

wherein R^(1b), R^(2b), R^(3b), and R^(4b) have the same meaning asdefined above. Preferably, the bond represented by a dashed and solidline represents a single bond and R^(1b) and R^(2b) are positioned as informula Ib or Id. Alternatively the bond represented by a dashed andsolid line represents a double bond.

In another embodiment, the invention provides compounds havingstructural Formula Ih, Ii, or Ij, and the stereoisomers, prodrugs,tautomers, racemates, salts, hydrates, or solvates thereof:

wherein R^(1b), R^(2b), R^(3b), and R^(4b) have the same meaning asdefined above. Preferably, the bond represented by a dashed and solidline represents a single bond and R^(1b) and R^(2b) are positioned as informula Ib or Id. Alternatively the bond represented by a dashed andsolid line represents a double bond.

Preferably, R^(3b) and R^(4b) have an anti-stereorelationship in thecompounds of formulas Ia to Ij defined herein.

Preferably, R^(1b) and R^(2b) are each independently selected fromhydrogen, hydroxyl, —NR¹⁰R¹¹, halogen, C₁₋₆alkyl, and C₁₋₆alkoxy,preferably R^(1b) and R^(2b) are both hydrogen, hydroxyl, halogen ormethoxy, or a combination of hydroxyl and methoxy in the compounds offormulas Ia to Ij defined herein.

In further preferred embodiments of the compounds of formulas Ia to Ijdefined herein, R^(3b) is selected from hydrogen, hydroxyl, —NR¹⁰R¹¹,halogen, C₁₋₆alkyl, and C₁₋₆alkoxy; preferably R^(3b) is hydroxyl,halogen, or methoxy.

In further preferred embodiments of the compounds of formulas Ia to Ijdefined herein, R^(4b) is selected from the group consisting ofhydrogen, hydroxyl, —NR¹⁰R¹¹, halogen, C₁₋₆alkyl, and C₁₋₆alkoxy;preferably R^(4b) is hydroxyl, halogen, or methoxy.

Further preferred embodiments of the invention are compounds having oneof the following structural formulas:

The invention further provides a pharmaceutical composition comprising acompound according to any one of the embodiments defined herein and apharmaceutically acceptable excipient. In another embodiment, saidpharmaceutical composition can comprise a further active pharmaceuticalingredient.

The invention further provides a compound according to any one of theembodiments defined herein, or the pharmaceutical composition as definedherein, for use as a medicament.

Preferably, for treating a disease by intervening at the level ofmicrotubule dynamics, more specifically by stabilizing the microtubules,more preferably for treating a proliferative disorder. In a furtherpreferred embodiment, said proliferative disorder is selected from thegroup comprising: neoplasia, dysplasia, tumor growth, or cancer.Preferred types of cancer are (advanced) breast cancer, prostate cancer,ovarian cancer, colorectal cancer, lung cancer, melanoma, or epidermoidcarcinoma, non-small cell lung cancer and Kaposi's sarcoma.

The invention further provides the use of the compounds or thepharmaceutical composition as defined herein for the manufacturing of amedicament for treating a disease caused by changes in microtubulestability, more preferably for treating a proliferative disorder. In afurther preferred embodiment, said proliferative disorder is selectedfrom the group comprising: neoplasia, dysplasia, tumor growth, orcancer. Preferred types of cancer are (advanced) breast cancer, prostatecancer, ovarian cancer, colorectal cancer, lung cancer, melanoma, orepidermoid carcinoma, non-small cell lung cancer and Kaposi's sarcoma.

The invention further provides a method of treating diseases as definedherein, in a subject needing such therapy, comprising administering atherapeutically effective amount of one or more of the compound(s) orthe pharmaceutical preparation defined herein to a patient in needthereof. More preferably said disease is a proliferative disorder. In afurther preferred embodiment, said proliferative disorder is selectedfrom the group comprising: neoplasia, dysplasia, tumor growth, orcancer. Preferred types of cancer are (advanced) breast cancer, prostatecancer, ovarian cancer, colorectal cancer, lung cancer, melanoma, orepidermoid carcinoma, non-small cell lung cancer and Kaposi's sarcoma.Optionally, said treatment of a proliferative disorder is performed incombination with any one of the therapies selected from the groupcomprising: surgery, chemotherapy, radiation therapy, immunotherapy,and/or gene therapy.

In certain embodiments of treatment, the compound or the pharmaceuticalpreparation as defined herein can be administered in combination withone or more additional active compounds, before, after or simultaneouslywith the administration of said compound or pharmaceutical composition.

In a preferred embodiment, the pharmaceutical composition as definedherein can be administered orally, for example in the form of pills,tablets, lacquered tablets, sugar-coated tablets, granules, hard andsoft gelatin capsules, aqueous, alcoholic or oily solutions, syrups,emulsions or suspensions; rectally, for example in the form ofsuppositories; parenterally, for example subcutaneously, intramuscularlyor intravenously in the form of solutions for injection or infusion;percutaneous or topically, for example in the form of ointments,tinctures, sprays or transdermal therapeutic systems; through inhalativeadministration in the form of e.g. nasal sprays or aerosol mixtures; orin the form of microcapsules, implants or rods.

In a preferred embodiment, the pharmaceutically acceptable carrierand/or additives can be selected from the group of: fillers,disintegrants, binders, lubricants, wetting agents, stabilizers,emulsifiers, dispersants, preservatives, sweeteners, colorants,flavorings, aromatizers, thickeners, diluents, buffer substances,solvents, solubilizers, agents for achieving a depot effect, salts foraltering the osmotic pressure, coating agents or antioxidants.

In a preferred embodiment, the subject is a mammal, e.g. human, equine,rabbit, mouse, rat, pig, sheep, cow or dog. Preferably the subject ishuman.

Taking into account the small size of the peloruside analog, we do notforesee pharmacodynamic problems in the systemic treatment of cancer.For “topical” treatment of ovarian cancer via the peritoneal cavity thesmall size may facilitate diffusion into the circulation, which reducesthe local retention of the drug at the site of tumor deposits in theperitoneum.

The invention further provides for a process for producing a compoundaccording to the general formula I, comprising the steps of:

-   -   (a) reacting a methyl ketone having structural Formula IIa with        an aldehyde having structural Formula IIb, through aldol        coupling, wherein P⁴ and P⁵ are protecting groups, and executing        suitable functional group interconversions, thereby obtaining a        compound having structural Formula II;    -   (b) protecting the functional groups at positions X³ and X⁴ in        the compound of Formula II where needed;    -   (c) reacting the compound of Formula II with a compound of the        Formula III in the presence of a suitable catalyst, giving rise        to a compound of the formula (IV),    -   (d) removing the protecting groups P¹ and P⁴ in the resulting        compound and esterifying the deprotected COOH with the C₁₅ OH        group, thereby obtaining the lactone having structural Formula        V; and    -   (e) deprotecting P⁵ in the compound having structural Formula V        and deprotecting any of the possibly protected X¹, X², X³, or X⁴        groups, if required after executing additional suitable        functional group interconversions, thereby obtaining a compound        having structural Formula I, generally following the reaction        scheme below:

-   -   wherein    -   P¹ is hydrogen or a carboxyl protecting group; for example        optionally substituted alkyl, preferably P¹ is methyl;    -   P² is selected from halogen, pseudohalogen, CF₃SO₃, OAc,        preferably P² is bromo, and simultaneously,    -   P³ is a trialkyltin, such as trimethyltin or tri-n-butyltin, or        is a boronic acid or boronic ester; or alternatively,    -   P² is a trialkyltin, such as trimethyltin or tri-n-butyltin, or        is a boronic acid or boronic ester and simultaneously,    -   P³ is selected from halogen, pseudohalogen, CF₃SO₃, OAc, (for        example, P³ is bromo);    -   P⁴ is selected from an orthogonally chosen protecting group,        preferably P⁴ is MPM (4-OMe-Bn);    -   P⁵ is a protecting group which can be orthogonally removed,        preferably selected from: TBS (tert-butyldimethylsilyl) or MEM        (2-methoxyethoxymethyl);    -   the suitable catalyst is typically a transition metal with        ligands. For example, said suitable catalyst can be        Pd₂(dba)₃.CHCl₃; and    -   X¹, X², X³, and X⁴ have the same meaning as that defined above,        and wherein, if present, their functional groups are suitably        protected in steps (b) and (c), functionally interconverted        where needed and deprotected accordingly in step (d).

BRIEF DESCRIPTION OF THE FIGURES

The present invention is illustrated by the following figures which areto be considered for illustrative purposes only and in no way limit theinvention to the embodiments disclosed therein:

FIG. 1: JC168 inhibits cell growth. MO4 cells were treated with dimethylsulfoxide (DMSO, solvent control), 0.001, 0.01, 0.1 or 1 μg/ml JC168 for4 days. After said 4 days of incubation the cells were fixed and stainedwith sulforhodamine B to determine cellular protein content. Excess dyewas removed by washing. Protein-bound dye was dissolved and opticaldensity was measured. Data represent optical density (OD) as apercentage of solvent control (mean and standard deviation, n=6).

FIG. 2: Comparison of JC168 and paclitaxel to inhibit cell growth. MO4cells were treated with dimethyl sulfoxide (DMSO, solvent control), 1μg/ml JC168 or 1 μg/ml paclitaxel for 4 days. After said 4 days ofincubation the cells were fixed and stained with sulforhodamine B todetermine cellular protein content. Excess dye was removed by washing.Protein-bound dye was dissolved and optical density was measured. Datarepresent optical density (OD) as a percentage of solvent control (meanand standard deviation, n=6).

FIG. 3: JC168 inhibits MTT conversion. MO4 cells were treated withdimethyl sulfoxide (DMSO, solvent control), 0.001, 0.01, 0.1 or 1 μg/mlJC168 for 4 days. Then,3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide was addedand the cultures were incubated to allow the cells to reduce the MTTinto formazan. The formazan was dissolved and optical density wasmeasured. Data represent optical density (OD) as a percentage of solventcontrol (mean and standard deviation, n=6).

FIG. 4: Comparison of JC168 and paclitaxel to inhibit MTT conversion.MO4 cells were treated with dimethyl sulfoxide (DMSO, solvent control),1 μg/ml JC168 or 1 μg/ml paclitaxel for 4 days. Then,3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide was addedand the cultures were incubated to allow the cells to reduce the MTTinto formazan. The formazan was dissolved and optical density wasmeasured. Data represent optical density (OD) as a percentage of solventcontrol (mean and standard deviation, n=6).

FIG. 5: JC168 inhibits invasion of MDA-MB231 GFP-LUC cells into collagentype I. Single cell suspensions of MDA-MB231 GFP-LUC cells wereincubated on top of a collagen type I gel in the absence (control) orpresence of DMSO (solvent control), 1 μg/ml paclitaxel, or 0.01, 0.1 or1 μg/ml JC168 for 24 hours. Invading cells were counted byphase-contrast microscopy as cells with extensions penetrating into thegel. (A) Invasion was calculated as the percentage of invading cells perhigh powered field (mean and standard deviation, n=10). Microphotographsof representative cultures of MDA-MB231 GFP-LUC cells treated with DMSO(B), 1 μg/ml JC168 (C), and 1 μg/ml paclitaxel (D) after 24 hours ofincubation on top of a collagen type I gel. Scalebar=100 μm.

FIG. 6 represents the ¹H NMR spectrum of a compound according to anembodiment of the present invention namely the compound designatedJC168.

FIG. 7 represents the ¹³C NMR spectrum of a compound according to anembodiment of the present invention namely the compound designatedJC168.

DETAILED DESCRIPTION OF THE INVENTION

As used herein, the singular forms “a”, “an”, and “the” include bothsingular and plural referents unless the context clearly dictatesotherwise. By way of example, “a compound” can refer to one or more thanone compound; “a composition” refers to one or more than onecomposition.

The terms “comprising”, “comprises” and “comprised of” as used hereinare synonymous with “including”, “includes” or “containing”, “contains”,and are inclusive or open-ended and do not exclude additional,non-recited members, elements or method steps. The term “comprising”also encompasses the closed-wording “consisting of” and “consistingessentially of”.

The term “about” as used herein when referring to a measurable valuesuch as a parameter, an amount, a temporal duration, and the like, ismeant to encompass variations of +/−20% or less, preferably +/−10% orless, more preferably +/−5% or less, even more preferably +/−1% or less,and still more preferably +/−0.1% or less from the specified value,insofar such variations are appropriate to perform in the disclosedinvention.

The expression “the corresponding R^(xa)” or “the corresponding R^(xb)group” needs to be seen in the light of formula “CR^(xa)R^(xb)” (e.g.CR^(1a)R^(1b), CR^(2a)R^(2b), CR^(3a)R^(3b), and CR^(4a)R^(4b)). Foreach R^(xa) group, there is a corresponding R^(xb) group in thesubstituent R^(xa)R^(xb) and vice versa. As an example, for each R^(1a)group, there is a corresponding R^(1b) group in the R^(1a)R^(1b)substituent, and vice versa.

All documents cited in the present specification are hereby incorporatedby reference in their entirety. In particular, the teachings of alldocuments herein specifically referred to are incorporated by reference.

The inventors have designed so-called “phenyl-pelorusides”, or“pelofens”, which are peloruside analogs, wherein the pyranose group hasbeen substituted by a phenyl group. The first compound synthesised wasdesignated “JC168” and had the following structural formula:

Because of the biological activity of JC168, a whole array of phenylanalogs can be synthesized to further explore the structure-activityrelationship (SAR) of the peloruside binding site. Because these phenylanalogs are structurally less complicated and thus more easilyaccessible than peloruside, the SAR of peloruside can be studied viathese analogs. The inventors therefore embarked on a strategy to designanalogs of said general pelofen compound, especially focussing onmodifying the substituents at positions C₂, C₃, C₁₁, and C₁₃. Saidpositions are called respectively X¹, X², X³ and X⁴ in general formulaI.

The term “compounds as defined herein” therefore comprises all compoundshaving the general Formula I:

-   -   wherein    -   X¹ is CR^(1a)R^(1b), X² is CR^(2a)R^(2b), X³ is CR^(3a)R^(3b),        X⁴ is CR^(4a)R^(4b), wherein    -   R^(1a), R^(2a), R^(3a), and R^(4a) are each independently        selected from hydrogen, hydroxyl, halogen, and a group selected        from —NR¹⁰R¹¹, C₁₋₆alkyl, C₂₋₆alkenyl, C₂₋₆alkynyl, C₆₋₁₀aryl,        C₆₋₁₀arylC₆alkyl and C₁₋₆alkoxy, each group independently being        optionally substituted with one or more substituent(s) each        independently selected from hydroxyl, halogen, C₁₋₆alkyl and        C₁₋₆alkoxy; and    -   R^(1b), R^(2b), R^(3b), and R^(4b) each independently are        selected from hydrogen, hydroxyl, halogen, and a group selected        from —NR¹⁰R¹¹, C₁₋₆alkyl, C₂₋₆alkenyl, C₂₋₆alkynyl, C₆₋₁₀aryl,        C₆₋₁₀arylC₁₋₆alkyl and C₁₋₆alkoxy, each group independently        being optionally substituted with one or more substituent(s)        each independently selected from hydroxyl, halogen, C₁₋₆alkyl        and C₁₋₆alkoxy; or    -   R^(1a) and R^(1b), or R^(2a) and R^(2b), or R^(3a) and R^(3b),        or R^(4a) and R^(4b) taken together represent an oxo (═O) group;    -   wherein the bond represented by a dashed and solid line        represents a single bond or a double bond and in case of a        double bond, R^(1b) and R^(2b) are absent and at least one of        R^(1a) and R^(2a) is not OH or —NR¹⁰R¹¹; and    -   R¹⁰ and R¹¹ are each independently selected from hydrogen and        C₁₋₆alkyl; and wherein:        -   when R^(1a), R^(2a), R^(3a) or R^(4a) is hydroxyl, the            corresponding R^(1b), R^(2b), R^(3b) or R^(4b) is not            hydroxyl, is not —NR¹⁰R¹¹, is not halogen, or is not            C₁₋₆alkoxy,        -   when R^(1a), R^(2a), R^(3a) or R^(4a) is —NR¹⁰R¹¹, the            corresponding R^(1b), R^(2b), R^(3b) or R^(4b) is not            hydroxyl, is not —NR¹⁰R¹¹, is not halogen, or is not            C₁₋₆alkoxy,        -   when R^(1a), R^(2a), R^(3a) or R^(4a) is halogen, the            corresponding R^(1b), R^(2b), R^(3b) or R^(4b) is not            hydroxyl, or is not —NR¹⁰R¹¹,        -   when R^(1b), R^(2b), R^(3b) or R^(4b) is hydroxyl, the            corresponding R^(1a), R^(2a), R^(3a) or R^(4a) is not            hydroxyl, is not —NR¹⁰R¹¹, is not halogen, or is not            C₁₋₆alkoxy, and        -   when R^(1b), R^(2b), R^(3b) or R^(4b) is —NR¹⁰R¹¹, the            corresponding R^(1a), R^(2a), R^(3a) or R^(4a) is not            hydroxyl, is not —NR¹⁰R¹¹, is not halogen, or is not            C₁₋₆alkoxy.        -   when R^(1b), R^(2b), R^(3b) or R^(4b) is halogen, the            corresponding R^(1a), R^(2a), R^(3a) or R^(4a) is not            hydroxyl, or is not —NR¹⁰R¹¹,

Also the stereoisomers, prodrugs, tautomers, racemates, salts, hydrates,or solvates of said compounds are falling within the definition of theterm “compounds as defined herein”.

In a preferred subset of compounds as defined herein, the C₂-C₃substituents are envisaged, meaning the structural formula I is onlychanged at said two positions. Positions C₁₁ and C₁₃ are as in theoriginal pelofen JC168, as depicted in formula (XI) below:

-   -   wherein        -   X¹ is CR^(1a)R^(1b), X² is CR^(2a)R^(2b), wherein        -   R^(1a) is selected from hydrogen, hydroxyl, halogen, and a            group selected from —NR¹⁰R¹¹, C₁₋₆alkyl, C₂₋₆alkenyl,            C₂₋₆alkynyl, C₆₋₁₀aryl, C₆₋₁₀arylC₁₋₆alkyl and C₁₋₆alkoxy,            each group independently being optionally substituted with            one or more substituent(s) each independently selected from            hydroxyl, halogen, C₁₋₆alkyl, and C₁₋₆alkoxy;        -   R^(2a) is selected from hydrogen, hydroxyl, halogen, and a            group selected from —NR¹⁰R¹¹, C₁₋₆alkyl, C₂₋₆alkenyl,            C₂₋₆alkynyl, C₆₋₁₀aryl, C₆₋₁₀arylC₁₋₆alkyl and C₁₋₆alkoxy,            each group independently being optionally substituted with            one or more substituent(s) each independently selected from            hydroxyl, halogen, C₁₋₆alkyl and C₁₋₆alkoxy;        -   R^(1b) is selected from hydrogen, hydroxyl, halogen and a            group selected from —NR¹⁰R¹¹, C₁₋₆alkyl, C₂₋₆alkenyl,            C₂₋₆alkynyl, C₆₋₁₀aryl, C₆₋₁₀arylC₁₋₆alkyl and C₁₋₆alkoxy,            each group independently being optionally substituted with            one or more substituent(s) each independently selected from            hydroxyl, halogen, C₁₋₆alkyl and C₁₋₆alkoxy;        -   R^(2b) is selected from hydrogen, hydroxyl, halogen and a            group selected from —NR¹⁰R¹¹, C₁₋₆alkyl, C₂₋₆alkenyl,            C₂₋₆alkynyl, C₆₋₁₀aryl, C₆₋₁₀arylC₁₋₆alkyl and C₁₋₆alkoxy,            each group independently being optionally substituted with            one or more substituent(s) each independently selected from            hydroxyl, halogen, C₁₋₆alkyl and C₁₋₆alkoxy; or wherein    -   R^(1a) and R^(1b), or R^(2a) and R^(2b), taken together        represent an oxo (═O) group;        -   wherein the bond represented by a dashed and solid line            represents a single bond or a double bond and in case of a            double bond, R^(1b) and R^(2b) are absent and at least one            of R^(1a) and R^(2a) is not OH or —NR¹⁰R¹¹; wherein        -   R¹⁰ and R¹¹ are each independently selected from hydrogen            and C₁₋₆alkyl; and wherein        -   when R^(1a) or R^(2a) is hydroxyl, the corresponding R^(1b)            or R^(2b) is not hydroxyl, is not —NR¹⁰R¹¹, is not halogen,            or is not C₁₋₆alkoxy,        -   when R^(1a) or R^(2a) is —NR¹⁰R¹¹, the corresponding R^(1b)            or R^(2b) is not hydroxyl, is not —NR¹⁰R¹¹, is not halogen,            or is not C₁₋₆alkoxy,        -   when R^(1a) or R^(2a) is halogen, the corresponding R^(1b)            or R^(2b) is not hydroxyl, or is not —NR¹⁰R¹¹,        -   when R^(1b) or R^(2b) is hydroxyl, the corresponding R^(1a)            or R^(2a) is not hydroxyl, is not —NR¹⁰R¹¹, is not halogen,            or is not C₁₋₆alkoxy, and        -   when R^(1b) or R^(2b) is —NR¹⁰R¹¹, the corresponding R^(1a)            or R^(2a) is not hydroxyl, is not —NR¹⁰R¹¹, is not halogen,            or is not C₁₋₆alkoxy.        -   when R^(1b) or R^(2b) is halogen, the corresponding R^(1a)            or R^(2a) is not hydroxyl, or is not —NR¹⁰R¹¹.

In preferred embodiments, the invention provides compounds of FormulaXIa:

wherein:

-   -   X¹ is CR^(1a)R^(1b), and X² is CR^(2a)R^(2b), wherein    -   R^(1a) and R^(2a) are both hydrogen;    -   R^(1b) and R^(2b) are each independently selected from hydrogen,        hydroxyl, —NR¹⁰R¹¹, halogen, C₁₋₆alkyl, and C₁₋₆alkoxy;        preferably wherein R^(1b) and R^(2b) are both hydrogen,        hydroxyl, halogen or methoxy, or a combination of hydroxyl and        methoxy; preferably, wherein R^(1b) and R^(2b) are halogen;        preferably, wherein R^(1b) is hydroxyl and R^(2b) is methoxy or        vice versa; preferably, wherein R^(1b) is halogen and R^(2b) is        methoxy, or vice versa; preferably, wherein R^(1b) is halogen        and R^(2b) is hydroxyl, or vice versa; preferably, wherein        R^(1b) and R^(2b) are methoxy; preferably, wherein R^(1b) and        R^(2b) are hydroxyl.

In further preferred embodiments, the invention provides compounds ofFormula XIb:

wherein:

-   -   X¹ is CR^(1a)R^(1b), and X² is CR^(2a)R^(2b), wherein    -   the bond between X¹ and X² is a double bond, and R^(1a) and        R^(2a) hence are absent;    -   R^(1b) and R^(2b) are each independently selected from hydrogen,        hydroxyl, —NR¹⁰R¹¹, halogen, C₁₋₆alkyl, and C₁₋₆alkoxy;        preferably wherein R^(1b) and R^(2b) are both hydrogen,        hydroxyl, halogen or methoxy, or a combination of hydroxyl and        methoxy; preferably, wherein R^(1b) and R^(2b) are halogen;        preferably, wherein R^(1b) is hydroxyl and R^(2b) is methoxy or        vice versa; preferably, wherein R^(1b) is halogen and R^(2b) is        methoxy, or vice versa; preferably, wherein R^(1b) is halogen        and R^(2b) is hydroxyl, or vice versa; preferably, wherein        R^(1b) and R^(2b) are methoxy; preferably, wherein R^(1b) and        R^(2b) are hydroxyl.

In another preferred subset of compounds as defined herein, the C₁₃substituents are envisaged, meaning the structural formula I is onlychanged at said position. Positions C₂, C₃, and C₁₁ are as in theoriginal pelofen JC168, as depicted in formula (XII) below:

-   -   wherein    -   X⁴ is CR^(4a)R^(4b), wherein        -   R^(4a) is selected from hydrogen, hydroxyl, halogen, and a            group selected from —NR¹⁰R¹¹, C₁₋₆alkyl, C₂₋₆alkenyl,            C₂₋₆alkynyl, C₆₋₁₀aryl, C₆₋₁₀arylC₁₋₆alkyl and C₁₋₆alkoxy,            each group independently being optionally substituted with            one or more substituent(s) each independently selected from            hydroxyl, halogen, C₁₋₆alkyl and C₁₋₆alkoxy; and        -   R^(4b) is selected from hydrogen, hydroxyl, halogen, and a            group selected from —NR¹⁰R¹¹, C₁₋₆alkyl, C₂₋₆alkenyl,            C₂₋₆alkynyl, C₆₋₁₀aryl, C₆₋₁₀arylC₁₋₆alkyl and C₁₋₆alkoxy,            each group independently being optionally substituted with            one or more substituent(s) each independently selected from            hydroxyl, halogen, C₁₋₆alkyl and C₁₋₆alkoxy; or wherein            R^(4a) and R^(4b) taken together represent an oxo (═O)            group; and wherein        -   R¹⁰ and R¹¹ are each independently selected from hydrogen            and C₁₋₆alkyl, and        -   when R^(4a) is hydroxyl, R^(4b) is not hydroxyl, is not            —NR¹⁰R¹¹, is not halogen, or is not C₁₋₆alkoxy,        -   when R^(4a) is —NR¹⁰R¹¹, R^(4b) is not hydroxyl, is not            —NR¹⁰R¹¹, is not halogen, or is not C₁₋₆alkoxy,        -   when R^(4a) is halogen, R^(4b) is not hydroxyl, or is not            —NR¹⁰R¹¹,        -   when R^(4b) is hydroxyl, R^(4a) is not hydroxyl, is not            —NR¹⁰R¹¹, is not halogen, or is not C₁₋₆alkoxy, and        -   when R^(4b) is —NR¹⁰R¹¹, R^(4a) is not hydroxyl, is not            —NR¹⁰R¹¹, is not halogen, or is not C₁₋₆alkoxy.        -   when R^(4b) is halogen, R^(4a) is not hydroxyl, or is not            —NR¹⁰R¹¹.

In preferred embodiments of the compounds of Formula XII, R^(4a) ishydrogen and R^(4b) is selected from the group consisting of: hydrogen,hydroxyl, halogen, or from a group selected from —NR¹⁰R¹¹, C₁₋₆alkyl,and C₁₋₆alkoxy, each independently optionally substituted with one ormore substituent(s) each independently selected from hydroxyl, halogen,C₁₋₆alkyl and C₁₋₆alkoxy. Preferably R^(4b) is hydroxyl, halogen, ormethoxy.

In further preferred embodiments of the compounds of Formula XII, R^(4a)and R^(4b) taken together represent an oxo (═O) group.

In another preferred subset of compounds as defined herein, the C₁₁substituents are envisaged, meaning the structural formula I is onlychanged at said position. Positions C₂, C₃, and C₁₃ are as in theoriginal pelofen JC168, as depicted in formula (XIII) below:

-   -   wherein    -   X³ is CR^(3a)R^(3b), wherein        -   R^(3a) is selected from hydrogen, hydroxyl, halogen, and a            group selected from —NR¹⁰R¹¹, C₁₋₆alkyl, C₂₋₆alkenyl,            C₂₋₆alkynyl, C₆₋₁₀aryl, C₆₋₁₀arylC₁₋₆alkyl and C₁₋₆alkoxy,            each group independently being optionally substituted with            one or more substituent(s) each independently selected from            hydroxyl, halogen, C₁₋₆alkyl and C₁₋₆alkoxy; and        -   R^(3b) is selected from hydrogen, hydroxyl, halogen, and a            group selected from —NR¹⁰R¹¹, C₁₋₆alkyl, C₂₋₆alkenyl,            C₂₋₆alkynyl, C₆₋₁₀aryl, C₆₋₁₀arylC₁₋₆alkyl and C₁₋₆alkoxy,            each group independently being optionally substituted with            one or more substituent(s) each independently selected from            hydroxyl, halogen, C₁₋₆alkyl and C₁₋₆alkoxy; or    -   R^(3a) and R^(3b) taken together represent an oxo (═O) group;    -   R¹⁰ and R¹¹ are each independently selected from hydrogen and        C₁₋₆alkyl, and        -   when R^(3a) is hydroxyl, R^(3b) is not hydroxyl, is not            —NR¹⁰R¹¹, is not halogen, or is not C₁₋₆alkoxy,        -   when R^(3a) is —NR¹⁰R¹¹, R^(3b) is not hydroxyl, is not            —NR¹⁰R¹¹, is not halogen, or is not C₁₋₆alkoxy,        -   when R^(3a) is halogen, R^(3b) is not hydroxyl, or is not            —NR¹⁰R¹¹,        -   when R^(3b) is hydroxyl, R^(3a) is not hydroxyl, is not            —NR¹⁰R¹¹, is not halogen, or is not C₁₋₆alkoxy, and        -   when R^(3b) is —NR¹⁰R¹¹, R^(3a) is not hydroxyl, is not            —NR¹⁰R¹¹, is not halogen, or is not C₁₋₆alkoxy.        -   when R^(3b) is halogen, R^(3a) is not hydroxyl, or is not            —NR¹⁰R¹¹

In further preferred embodiments of the compounds defined by Formula(XIII), R^(3a) is hydrogen and R^(3b) is selected from the groupconsisting of: hydrogen, hydroxyl, halogen, or from a group selectedfrom: —NR¹⁰R¹¹, C₁₋₆alkyl, and C₁₋₆alkoxy, each independently optionallysubstituted with one or more substituent(s) each independently selectedfrom hydroxyl, halogen, C₁₋₆alkyl and C₁₋₆alkoxy. Preferably R^(3b) ishydroxyl, halogen, or methoxy.

In further preferred embodiments of the compounds defined by FormulaXIII, R^(3a) and R^(3b) taken together represent an oxo (═O) group.

In yet another subset of compounds, as defined herein, both the C₁₁ andC₁₃ substituents are envisaged, meaning the structural formula I ischanged at positions C₁₁ and C₁₃. Positions C₂, and C₃, are as in theoriginal pelofen JC168, as depicted in formula (XIV) below:

-   -   wherein    -   X³ is CR^(3a)R^(3b), X⁴ is CR^(4a)R^(4b), wherein        -   R^(3a) is selected from hydrogen, hydroxyl, halogen, and a            group selected from —NR¹⁰R¹¹, C₁₋₆alkyl, C₂₋₆alkenyl,            C₂₋₆alkynyl, C₆₋₁₀aryl, C₆₋₁₀arylC₁₋₆alkyl and C₁₋₆alkoxy,            each group independently being optionally substituted with            one or more substituent(s) each independently selected from            hydroxyl, halogen, C₁₋₆alkyl and C₁₋₆alkoxy;        -   R^(4a) is selected from hydrogen, hydroxyl, halogen, and a            group selected from —NR¹⁰R¹¹, C₁₋₆alkyl, C₂₋₆alkenyl,            C₂₋₆alkynyl, C₆₋₁₀aryl, C₆₋₁₀arylC₁₋₆alkyl and C₁₋₆alkoxy,            each group independently being optionally substituted with            one or more substituent(s) each independently selected from            hydroxyl, halogen, C₁₋₆alkyl and C₁₋₆alkoxy; and        -   R^(3b) is selected from hydrogen, hydroxyl, halogen, and a            group selected from —NR¹⁰R¹¹, C₁₋₆alkyl, C₂₋₆alkenyl,            C₂₋₆alkynyl, C₆₋₁₀aryl, C₆₋₁₀arylC₁₋₆alkyl and C₁₋₆alkoxy,            each group independently being optionally substituted with            one or more substituent(s) each independently selected from            hydroxyl, halogen, C₁₋₆alkyl and C₁₋₆alkoxy;        -   R^(4b) is selected from hydrogen, hydroxyl, halogen, and a            group selected from —NR¹⁰R¹¹, C₁₋₆alkyl, C₂₋₆alkenyl,            C₂₋₆alkynyl, C₆₋₁₀aryl, C₆₋₁₀arylC₁₋₆alkyl and C₁₋₆alkoxy,            each group independently being optionally substituted with            one or more substituent(s) each independently selected from            hydroxyl, halogen, C₁₋₆alkyl and C₁₋₆alkoxy; or wherein        -   R^(3a) and R^(3b), or R^(4a) and R^(4b) taken together            represent an oxo (═O) group; and wherein        -   R¹⁰ and R¹¹ are each independently selected from hydrogen            and C₁₋₆alkyl, and        -   when R^(3a) or R^(4a) is hydroxyl, the corresponding R^(3b)            or R^(4b) is not hydroxyl, is not —NR¹⁰R¹¹, is not halogen,            or is not C₁₋₆alkoxy,        -   when R^(3a) or R^(4a) is —NR¹⁰R¹¹, the corresponding R^(3b)            or R^(4b) is not hydroxyl, is not —NR¹⁰R¹¹, is not halogen,            or is not C₁₋₆alkoxy,        -   when R^(3a) or R^(4a) is halogen, the corresponding R^(3b)            or R^(4b) is not hydroxyl, or is not —NR¹⁰R¹¹,        -   when R^(3b) or R^(4b) is hydroxyl, the corresponding R^(3a)            or R^(4a) is not hydroxyl, is not —NR¹⁰R¹¹, is not halogen,            or is not C₁₋₆alkoxy, and        -   when R^(3b) or R^(4b) is —NR¹⁰R¹¹, the corresponding R^(3a)            or R^(4a) is not hydroxyl, is not —NR¹⁰R¹¹, is not halogen,            or is not C₁₋₆alkoxy.        -   when R^(3b) or R^(4b) is halogen, the corresponding R^(3a)            or R^(4a) is not hydroxyl, or is not —NR¹⁰R¹¹

In further preferred embodiments of the compounds defined by Formula(XIV),

-   -   R^(3a) and R^(4a) are both hydrogen,    -   R^(3b) is selected from the group consisting of: hydrogen,        hydroxyl, halogen, or from a group selected from: —NR¹⁰R¹¹,        C₁₋₆alkyl, and C₁₋₆alkoxy, each independently optionally        substituted with one or more substituent(s) each independently        selected from hydroxyl, halogen, C₁₋₆alkyl and C₁₋₆alkoxy,        and/or    -   R^(4b) is selected from the group consisting of: hydrogen,        hydroxyl, halogen, or from a group selected from —NR¹⁰R¹¹,        C₁₋₆alkyl, and C₁₋₆alkoxy, each independently optionally        substituted with one or more substituent(s) each independently        selected from hydroxyl, halogen, C₁₋₆alkyl and C₁₋₆alkoxy.        Preferably R^(3b) is hydroxyl and R^(4b) is methoxy or vice        versa. Preferably, R^(3b) is hydroxyl and R^(4b) is halogen or        vice versa. Preferably, R^(3b) is halogen and R^(4b) is methoxy        or vice versa. Preferably, both R^(3b) and R^(4b) are hydroxyl,        both R^(3b) and R^(4b) are halogen, or both R^(3b) and R^(4b)        are methoxy.

In yet another subset of compounds, as defined herein, all four C₂, C₃,C₁₁ and C₁₃ substituents are envisaged, meaning the structural formula Iis changed at positions C₂, C₃, C₁₁ and C₁₃:

-   -   wherein        -   X¹ is CR^(1a)R^(1b), X² is CR^(2a)R^(2b), X³ is            CR^(3a)R^(3b), X⁴ is CR^(4a)R^(4b), and wherein:        -   R^(1a) is selected from hydrogen, hydroxyl, halogen, and a            group selected from —NR¹⁰R¹¹, C₁₋₆alkyl, C₂₋₆alkenyl,            C₂₋₆alkynyl, C₆₋₁₀aryl, C₆₋₁₀arylC₁₋₆alkyl and C₁₋₆alkoxy,            each group being independently optionally substituted with            one or more substituent(s) each independently selected from            hydroxyl, halogen, C₁₋₆alkyl and C₁₋₆alkoxy;        -   R^(2a) is selected from hydrogen, hydroxyl, halogen, and a            group selected from —NR¹⁰R¹¹, C₁₋₆alkyl, C₂₋₆alkenyl,            C₂₋₆alkynyl, C₆₋₁₀aryl, C₆₋₁₀arylC₁₋₆alkyl and C₁₋₆alkoxy,            each group being independently optionally substituted with            one or more substituent(s) each independently selected from            hydroxyl, halogen, C₁₋₆alkyl and C₁₋₆alkoxy,        -   R^(3a) is selected from hydrogen, hydroxyl, halogen, and a            group selected from —NR¹⁰R¹¹, C₁₋₆alkyl, C₂₋₆alkenyl,            C₂₋₆alkynyl, C₆₋₁₀aryl, C₆₋₁₀arylC₁₋₆alkyl and C₁₋₆alkoxy,            each group being independently optionally substituted with            one or more substituent(s) each independently selected from            hydroxyl, halogen, C₁₋₆alkyl and C₁₋₆alkoxy        -   R^(4a) is selected from hydrogen, hydroxyl, halogen, and a            group selected from —NR¹⁰R¹¹, C₁₋₆alkyl, C₂₋₆alkenyl,            C₂₋₆alkynyl, C₆₋₁₀aryl, C₆₋₁₀arylC₁₋₆alkyl and C₁₋₆alkoxy,            each group being independently optionally substituted with            one or more substituent(s) each independently selected from            hydroxyl, halogen, C₁₋₆alkyl and C₁₋₆alkoxy; and wherein:        -   R^(1b) is selected from hydrogen, hydroxyl, halogen, and a            group selected from —NR¹⁰R¹¹, C₁₋₆alkyl, C₂₋₆alkenyl,            C₂₋₆alkynyl, C₆₋₁₀aryl, C₆₋₁₀arylC₁₋₆alkyl and C₁₋₆alkoxy,            each group being independently optionally substituted with            one or more substituent(s) each independently selected from            hydroxyl, halogen, C₁₋₆alkyl and C₁₋₆alkoxy,        -   R^(2b) is selected from hydrogen, hydroxyl, halogen, and a            group selected from —NR¹⁰R¹¹, C₁₋₆alkyl, C₂₋₆alkenyl,            C₂₋₆alkynyl, C₆₋₁₀aryl, C₆₋₁₀arylC₁₋₆alkyl and C₁₋₆alkoxy,            each group being independently optionally substituted with            one or more substituent(s) each independently selected from            hydroxyl, halogen, C₁₋₆alkyl and C₁₋₆alkoxy,        -   R^(3b) is selected from hydrogen, hydroxyl, halogen, and a            group selected from —NR¹⁰R¹¹, C₁₋₆alkyl, C₂₋₆alkenyl,            C₂₋₆alkynyl, C₆₋₁₀aryl, C₆₋₁₀arylC₁₋₆alkyl and C₁₋₆alkoxy,            each group being independently optionally substituted with            one or more substituent(s) each independently selected from            hydroxyl, halogen, C₁₋₆alkyl and C₁₋₆alkoxy,        -   R^(4b) is selected from hydrogen, hydroxyl, halogen, and a            group selected from —NR¹⁰R¹¹, C₁₋₆alkyl, C₂₋₆alkenyl,            C₂₋₆alkynyl, C₆₋₁₀aryl, C₆₋₁₀arylC₁₋₆alkyl and C₁₋₆alkoxy,            each group independently being independently optionally            substituted with one or more substituent(s) each            independently selected from hydroxyl, halogen, C₁₋₆alkyl and            C₁₋₆alkoxy;    -   or wherein R^(1a) and R^(1b), or R^(2a) and R^(2b), or R^(3a)        and R^(3b), or R^(4a) and R^(4b) taken together represent an oxo        (═O) group; and wherein the bond represented by a dashed and        solid line represents a single bond or a double bond and wherein        in case of a double bond, R^(1b) and R^(2b) are absent and at        least one of R^(1a) and R^(2a) is not OH or —NR¹⁰R¹¹; and        wherein:        -   R¹⁰ and R¹¹ are each independently selected from hydrogen            and C₁₋₆alkyl; and wherein:        -   when R^(1a), R^(2a), R^(3a) or R^(4a) is hydroxyl, the            corresponding R^(1b), R^(2b), R^(3b) or R^(4b) is not            hydroxyl, is not —NR¹⁰R¹¹, is not halogen, or is not            C₁₋₆alkoxy,        -   when R^(1a), R^(2a), R^(3a) or R^(4a) is —NR¹⁰R¹¹, the            corresponding R^(1b), R^(2b), R^(3b) or R^(4b) is not            hydroxyl, is not —NR¹⁰R¹¹, is not halogen, or is not            C₁₋₆alkoxy,        -   when R^(1a), R^(2a), R^(3a) or R^(4a) is halogen, the            corresponding R^(1b), R^(2b), R^(3b) or R^(4b) is not            hydroxyl, or is not —NR¹⁰R¹¹,        -   when R^(1b), R^(2b), R^(3b) or R^(4b) is hydroxyl, the            corresponding R^(1a), R^(2a), R^(3a) or R^(4a) is not            hydroxyl, is not —NR¹⁰R¹¹, is not halogen, or is not            C₁₋₆alkoxy, and        -   when R^(1b), R^(2b), R^(3b) or R^(4b) is —NR¹⁰R¹¹, the            corresponding R^(1a), R^(2a), R^(3a) or R^(4a) is not            hydroxyl, is not —NR¹⁰R¹¹, is not halogen, or is not            C₁₋₆alkoxy.        -   when R^(1b), R^(2b), R^(3b) or R^(4b) is halogen, the            corresponding R^(1a), R^(2a), R^(3a) or R^(4a) is not            hydroxyl, or is not —NR¹⁰R¹¹.

In a preferred embodiment, the invention provides compounds havingstructural Formula Ia, and the stereoisomers, prodrugs, tautomers,racemates, salts, hydrates, or solvates thereof:

wherein R^(1b), R^(2b), R^(3b), and R^(4b) have the same meaning asdefined above.

In another preferred embodiment, the invention provides compounds havingstructural Formula Ib, Ic or Id, and the stereoisomers, prodrugs,tautomers, racemates, salts, hydrates, or solvates thereof:

wherein R^(1b), R^(2b), R^(3b), and R^(4b) have the same meaning asdefined above. The double bond in structure Ic can have the Z- orE-geometry.

In another preferred embodiment, the invention provides compounds havingstructural Formula Ie, If, or Ig, and the stereoisomers, prodrugs,tautomers, racemates, salts, hydrates, or solvates thereof:

wherein R^(1b), R^(2b), R^(3b), and R^(4b) have the same meaning asdefined above. Preferably, the bond represented by a dashed and solidline represents a single bond and R^(1b) and R^(2b) are positioned as informula Ib or Id. Alternatively the bond represented by a dashed andsolid line represents a double bond.

In another embodiment, the invention provides compounds havingstructural Formula Ih, Ii, or Ij, and the stereoisomers, prodrugs,tautomers, racemates, salts, hydrates, or solvates thereof:

wherein R^(1b), R^(2b), R^(3b), and R^(4b) have the same meaning asdefined above. Preferably, the bond represented by a dashed and solidline represents a single bond and R^(1b) and R^(2b) are positioned as informula Ib or Id. Alternatively the bond represented by a dashed andsolid line represents a double bond.

Preferably, R^(3b) and R^(4b) have an anti-stereorelationship in thecompounds defined herein.

Preferably, R^(1b) and R^(2b) are each independently selected fromhydrogen, hydroxyl, —NR¹⁰R¹¹, halogen, C₁₋₆alkyl, and C₁₋₆alkoxy,preferably R^(1b) and R^(2b) are both hydrogen, hydroxyl, halogen ormethoxy, or a combination of hydroxyl and methoxy in the compoundsdefined herein.

In further preferred embodiments of the compounds defined herein, R^(3b)is selected from hydrogen, hydroxyl, —NR¹⁰R¹¹, halogen, C₁₋₆alkyl, andC₁₋₆alkoxy; preferably R^(3b) is hydroxyl, halogen, or methoxy.

In further preferred embodiments of the compounds defined herein, R^(4b)is selected from the group consisting of hydrogen, hydroxyl, —NR¹⁰R¹¹,halogen, C₁₋₆alkyl, and C₁₋₆alkoxy; preferably R^(4b) is hydroxyl,halogen, or methoxy.

Further preferred embodiments of the invention are compounds having oneof the following structural formulas:

In the compounds defined herein, the term “halo” or “halogen” as a groupor part of a group is generic for fluoro, chloro, bromo, or iodo.

In the compounds defined herein, the term “oxo” as used herein refers tothe group ═O.

In the compounds defined herein, the term “hydroxyl” or “hydroxy” asused herein refers to the group —OH.

The term “alkyl”, as a group or part of a group, refers to a hydrocarbylgroup of Formula C_(n)H_(2n+1) wherein n is a number of at least 1.Alkyl groups may be linear, or branched and may be substituted asindicated herein. Generally, the alkyl groups comprise from 1 to 6carbon atoms, more preferably 1, 2, 3, 4, 5, or 6 carbon atoms. When asubscript is used herein following a carbon atom, the subscript refersto the number of carbon atoms that the named group may contain. Forexample, the term “C₁₋₆alkyl”, as a group or part of a group, refers toa hydrocarbyl group of Formula C_(n)H_(2n+1) wherein n is a numberranging from 1 to 6. Thus, for example, C₁₋₂₀alkyl groups include alllinear, or branched alkyl groups having 1 to 6 carbon atoms, and thusincludes for example methyl, ethyl, n-propyl, i-propyl, 2-methyl-ethyl,butyl and its isomers (e.g. n-butyl, i-butyl and t-butyl); pentyl andits isomers, hexyl and its isomers. For example, C₁₋₆ alkyl includes alllinear, or branched alkyl groups having 1 to 6 carbon atoms, and thusincludes for example methyl, ethyl, n-propyl, i-propyl, 2-methyl-ethyl,butyl and its isomers (e.g. n-butyl, i-butyl and t-butyl); pentyl andits isomers, hexyl and its isomers. When the suffix “ene” is used inconjunction with an alkyl group, i.e. “alkylene”, this is intended tomean the alkyl group as defined herein having two single bonds as pointsof attachment to other groups. Non-limiting examples of alkylene groupsincludes methylene, ethylene, methylmethylene, propylene, ethylethylene,and 1,2-dimethylethylene. Similarly, where alkenyl groups as definedherein and alkynyl groups as defined herein, respectively, are divalentgroups having single bonds for attachment to two other groups, they aretermed “alkenylene” and “alkynylene” respectively.

In the compounds defined herein, the term “C₂₋₆alkenyl” as a group orpart of a group, refers to an unsaturated hydrocarbyl group, which maybe linear, branched or cyclic, comprising one or more carbon-carbondouble bonds. Alkenyl groups thus preferably comprise between 2 and 6carbon atoms. Examples of alkenyl groups are ethenyl, 2-propenyl,2-butenyl, 3-butenyl, 2-pentenyl and its isomers, 2-hexenyl and itsisomers and the like.

In the compounds defined herein, the term “C₂₋₆alkynyl” as a group orpart of a group, refers to a class of monovalent unsaturated hydrocarbylgroups, wherein the unsaturation arises from the presence of one or morecarbon-carbon triple bonds. Alkynyl groups thus preferably comprisebetween 2 and 6 carbon atoms. Non limiting examples of alkynyl groupsare ethynyl, 2-propynyl, 2-butynyl, 3-butynyl, 2-pentynyl and itsisomers, 2-hexynyl and its isomers and the like.

In the compounds defined herein, the term “C₁₋₆alkoxy”, as a group orpart of a group, refers to a group having the Formula —OR^(a) whereinR^(a) is C₁₋₆alkyl as defined herein above. Non-limiting examples ofsuitable C₁₋₆alkoxy include methoxy, ethoxy, propoxy, isopropoxy,butoxy, isobutoxy, sec-butoxy, tert-butoxy, pentyloxy and hexyloxy.

In the compounds defined herein, the term “C₆₋₁₀aryl”, as a group orpart of a group, refers to a polyunsaturated, aromatic hydrocarbyl grouphaving a single ring (i.e. phenyl) or multiple aromatic rings fusedtogether (e.g. naphthalene), or linked covalently, typically containing6 to 10 atoms; wherein at least one ring is aromatic. The aromatic ringmay optionally include one to two additional rings (either cycloalkyl,heterocyclyl or heteroaryl) fused thereto. Examples of suitable arylinclude C6-10aryl, more preferably C6-8aryl. Non-limiting examplescomprise phenyl, biphenylyl, biphenylenyl, or 1- or 2-naphthanelyl; 5-or 6-tetralinyl, 1-, 2-, 3-, 4-, 5-, 6-, 7- or 8-azulenyl, 4-, 5-, 6 or7-indenyl, 4- or 5-indanyl, 5-, 6-, 7- or 8-tetrahydronaphthyl,1,2,3,4-tetrahydronaphthyl, and 1,4-dihydronaphthyl. When the suffix“ene” is used in conjunction with an aryl group, this is intended tomean the aryl group as defined herein having two single bonds as pointsof attachment to other groups, such as phenylene, biphenylylene,naphthylene, indenylene, and the like. Where a carbon atom in an arylgroup is replaced with a heteroatom, the resultant ring is referred toherein as a heteroaryl ring.

In the compounds defined herein, the term “C₆₋₁₀arylC₁₋₆alkyl”, as agroup or part of a group, means a C₁₋₆alkyl as defined herein, whereinat least one hydrogen atom is replaced by at least one C₆₋₁₀aryl asdefined herein. Non-limiting examples of C₆₋₁₀arylC₁₋₆alkyl groupinclude benzyl, phenethyl, dibenzylmethyl, methylphenylmethyl,3-(2-naphthyl)-butyl, and the like.

The term “proliferative disease or disorder” is meant to include allneoplastic cell growth and proliferation, whether malignant or benign,including all transformed cells and tissues and all cancerous cells andtissues. Proliferative diseases or disorders include, but are notlimited to, premalignant or precancerous lesions, abnormal cell growths,benign tumours, malignant tumours, and cancer.

Additional examples of proliferative diseases and/or disorders include,but are not limited to neoplasms, whether benign or malignant, locatedin the prostate, colon, abdomen, bone, breast, digestive system, liver,pancreas, peritoneum, endocrine glands (adrenal, parathyroid, pituitary,testicles, ovary, thymus, thyroid), eye, head and neck, nervous (centraland peripheral), lymphatic system, pelvic, skin, soft tissue, spleen,thoracic, and/or urogenital tract. In a preferred embodiment, theproliferative disorder involves tumour.

As used herein, the terms “tumour” or “tumour tissue” refer to anabnormal mass of tissue that results from excessive cell division. Atumour or tumour tissue comprises “tumour cells” which are neoplasticcells with abnormal growth properties and no useful bodily function.Tumours, tumour tissue and tumour cells may be benign or malignant. Atumour or tumour tissue may also comprise “tumour-associated non-tumourcells”, e.g., vascular cells which form blood vessels to supply thetumour or tumour tissue. Non-tumour cells may be induced to replicateand develop by tumour cells, for example, the induction of angiogenesisin a tumour or tumour tissue. In another preferred embodiment, theproliferative disorder involves malignancy or cancer.

As used herein, the term “malignancy” refers to a non-benign tumour or acancer. As used herein, the term “cancer” connotes a type ofproliferative disease which includes a malignancy characterized byderegulated or uncontrolled cell growth. Examples of cancer include, butare not limited to, carcinoma, lymphoma, blastoma, sarcoma, and leukemiaor lymphoid malignancies. More particular examples of such cancers arenoted below and include: squamous cell cancer (e.g., epithelial squamouscell cancer), lung cancer including small-cell lung cancer, non-smallcell lung cancer, adenocarcinoma of the lung, squamous carcinoma of thelung and large cell carcinoma of the lung, cancer of the peritoneum,hepatocellular cancer, gastric or stomach cancer includinggastrointestinal cancer, pancreatic cancer, glioblastoma, cervicalcancer, ovarian cancer, Kaposi's sarcoma, liver cancer, bladder cancer,hepatoma, breast cancer, colon cancer, rectal cancer, colorectal cancer,endometrial cancer or uterine carcinoma, salivary gland carcinoma,kidney or renal cancer, prostate cancer, vulval cancer, thyroid cancer,hepatic carcinoma, anal carcinoma, penile carcinoma, as well as CNScancer, melanoma, head and neck cancer, bone cancer, bone marrow cancer,duodenum cancer, oesophageal cancer, thyroid cancer, hematologicalcancer. The term “cancer” includes primary malignant cells or tumours(e.g., those whose cells have not migrated to sites in the subject'sbody other than the site of the original malignancy or tumour) andsecondary malignant cells or tumours (e.g., those arising frommetastasis, the migration of malignant cells or tumour cells tosecondary sites that are different from the site of the originaltumour).

In a further embodiment, the proliferative disorder is a premalignantcondition. Premalignant conditions are known or suspected of precedingprogression to neoplasia or cancer, in particular, where non-neoplasticcell growth consisting of hyperplasia, metaplasia, or most particularly,dysplasia has occurred (for review of such abnormal growth conditions,see Robbins and Angell 1976 (Basic Pathology, 2d Ed., W. B. SaundersCo., Philadelphia, pp. 68-79).

“Hyperplasia” is a form of controlled cell proliferation, involving anincrease in cell number in a tissue or organ, without significantalteration in structure or function. Hyperplastic disorders which can betreated by the method of the invention include, but are not limited to,angiofollicular mediastinal lymph node hyperplasia, angiolymphoidhyperplasia with eosinophilia, atypical melanocytic hyperplasia, basalcell hyperplasia, benign giant lymph node hyperplasia, cementumhyperplasia, congenital adrenal hyperplasia, congenital sebaceoushyperplasia, cystic hyperplasia, cystic hyperplasia of the breast,denture hyperplasia, ductal hyperplasia, endometrial hyperplasia,fibromuscular hyperplasia, focal epithelial hyperplasia, gingivalhyperplasia, inflammatory fibrous hyperplasia, inflammatory papillaryhyperplasia, intravascular papillary endothelial hyperplasia, nodularhyperplasia of prostate, nodular regenerative hyperplasia,pseudoepitheliomatous hyperplasia, senile sebaceous hyperplasia, andverrucous hyperplasia.

“Metaplasia” is a form of controlled cell growth in which one type ofadult or fully differentiated cell substitutes for another type of adultcell. Metaplastic disorders which can be treated by the method of theinvention include, but are not limited to, agnogenic myeloid metaplasia,apocrine metaplasia, atypical metaplasia, autoparenchymatous metaplasia,connective tissue metaplasia, epithelial metaplasia, intestinalmetaplasia, metaplastic anemia, metaplastic ossification, metaplasticpolyps, myeloid metaplasia, primary myeloid metaplasia, secondarymyeloid metaplasia, squamous metaplasia, squamous metaplasia of amnion,and symptomatic myeloid metaplasia.

“Dysplasia” is frequently a forerunner of cancer, and is found mainly inthe epithelia; it is the most disorderly form of non-neoplastic cellgrowth, involving a loss in individual cell uniformity and in thearchitectural orientation of cells. Dysplastic cells often haveabnormally large, deeply stained nuclei, and exhibit pleomorphism.Dysplasia characteristically occurs where there exists chronicirritation or inflammation. Dysplastic disorders which can be treated bythe method of the invention include, but are not limited to, anhidroticectodermal dysplasia, anterofacial dysplasia, asphyxiating thoracicdysplasia, atriodigital dysplasia, bronchopulmonary dysplasia, cerebraldysplasia, cervical dysplasia, chondroectodermal dysplasia,cleidocranial dysplasia, congenital ectodermal dysplasia,craniodiaphysial dysplasia, craniocarpotarsal dysplasia,craniometaphysial dysplasia, dentin dysplasia, diaphysial dysplasia,ectodermal dysplasia, enamel dysplasia, encephalo-ophthalmic dysplasia,dysplasia epiphysialis hemimelia, dysplasia epiphysialis multiplex,dysplasia epiphysialis punctata, epithelial dysplasia,faciodigitogenital dysplasia, familial fibrous dysplasia of jaws,familial white folded dysplasia, fibromuscular dysplasia, fibrousdysplasia of bone, florid osseous dysplasia, hereditary renal-retinaldysplasia, hidrotic ectodermal dysplasia, hypohidrotic ectodermaldysplasia, lymphopenic thymic dysplasia, mammary dysplasia,mandibulofacial dysplasia, metaphysial dysplasia, Mondini dysplasia,monostotic fibrous dysplasia, mucoepithelial dysplasia, multipleepiphysial dysplasia, oculoauriculovertebral dysplasia,oculodentodigital dysplasia, oculovertebral dysplasia, odontogenicdysplasia, ophthalmomandibulomelic dysplasia, periapical cementaldysplasia, polyostotic fibrous dysplasia, pseudoachondroplasticspondyloepiphysial dysplasia, retinal dysplasia, septo-optic dysplasia,spondyloepiphysial dysplasia, and ventriculoradial dysplasia. Additionalpre-neoplastic disorders include, but are not limited to, benigndysproliferative disorders (e.g., benign tumours, fibrocysticconditions, tissue hypertrophy, intestinal polyps, colon polyps, andoesophageal dysplasia), leukoplakia, keratoses, Bowen's disease,Farmer's Skin, solar cheilitis, and solar keratosis.

In preferred embodiments, the proliferative disorder is chosen fromglioma, preferably glioblastoma; prostate cancer; non-small-cell lungcancer (NSCLC); melanoma, head and neck cancer, pancreas cancer or coloncancer. By showing the anti-proliferative effect of the compounds of theinvention on cell-lines derived from each of these cancer-types, theinventors realised that the above cancer types can particularly benefitfrom the methods and agents of the invention.

As used herein, the term “glioma” refers to its art-recognisedconnotation. By virtue of further illustration and not limitation, theterm “glioma” refers to a tumour originating in the neuroglia of thebrain or spinal cord. Gliomas can be derived from glial cell types, suchas, e.g., astrocytes and oligodendrocytes, thus gliomas includeastrocytomas and oligodendrogliomas, as well as anaplastic gliomas,glioblastomas, and ependymomas. Astrocytomas and ependymomas can occurin all areas of the brain and spinal cord in both children and adults.Oligodendrogliomas typically occur in the cerebral hemispheres ofadults. Malignant astrocytic gliomas are associated with the worstprognoses because of their ability to infiltrate diffusely into thenormal brain parenchyma and include World Health Organization (WHO)grades II, III and grade IV tumors.

The present invention also provides methods of treating proliferativedisorders in a subject needing such therapy, comprising administering atherapeutically effective amount of the compound or the pharmaceuticalcomposition as defined herein.

As used herein, the terms “treat” or “treatment” refer to boththerapeutic treatment and prophylactic or preventative measures, whereinthe objective is to prevent or slow down (lessen) an undesiredphysiological change or disorder, such as the development or spread ofproliferative disease, e.g., cancer. Beneficial or desired clinicalresults include, but are not limited to, alleviation of symptoms,diminishment of extent of disease, stabilised (i.e., not worsening)state of disease, delay or slowing of disease progression, ameliorationor palliation of the disease state, and remission (whether partial ortotal), whether detectable or undetectable. “Treatment” can also meanprolonging survival as compared to expected survival if not receivingtreatment.

As used herein, a phrase such as “a subject in need of treatment”includes subjects, such as mammalian subjects, that would benefit fromtreatment of a given condition, preferably a proliferative disease, suchas, e.g., cancer, e.g., as above.

Such subjects will typically include, without limitation, those thathave been diagnosed with the condition, preferably a proliferativedisease, e.g., cancer, those prone to have or develop the said conditionand/or those in whom the condition is to be prevented.

The term “therapeutically effective amount” refers to an amount of acompound or pharmaceutical composition as defined herein effective totreat a disease or disorder in a subject, i.e., to obtain a desiredlocal or systemic effect and performance. By means of example and notlimitation, in the case of proliferative disease, e.g., cancer,therapeutically effective amount of a drug may reduce the number ofcancer cells; reduce the tumour size; inhibit (i.e., slow to some extentand preferably stop) cancer cell infiltration into peripheral organs;inhibit (i.e., slow to some extent and preferably stop) tumourmetastasis; inhibit, to some extent, tumour growth; enhance efficacy ofanother cancer therapy; and/or relieve to some extent one or more of thesymptoms associated with the cancer. To the extent the drug may preventgrowth and/or kill existing cancer cells, it may be cytostatic and/orcytotoxic. For cancer therapy, efficacy can, for example, be measured byassessing the time to disease progression (TTP) and/or determining theresponse rate (RR). The term thus refers to the quantity of compound orpharmaceutical composition that elicits the biological or medicinalresponse in a tissue, system, animal, or human that is being sought by aresearcher, veterinarian, medical doctor or other clinician, whichincludes alleviation of the symptoms of the cancer being treated. Inparticular, these terms refer to the quantity of compound orpharmaceutical composition according to the invention which is necessaryto prevent, cure, ameliorate, or at least minimize the clinicalimpairment, symptoms, or complications associated with cancer in eithera single or multiple doses.

The compound or the pharmaceutical composition as defined herein may beused alone or in combination with any of the cancer therapies selectedfrom the group comprising chemotherapy, radiation therapy,immunotherapy, and/or gene therapy. As used herein the term “cancertherapy” is meant to encompass radiation therapy, chemotherapy,immunotherapy, gene-based therapy, surgery, as well as combinationsthereof.

In another preferred embodiment the compound or the pharmaceuticalcomposition as defined herein may be used alone or in combination withone or more active compounds that are suitable in the treatment ofcancer. The term “active compound” refers to a compound other than theagents of the invention which is used to treat cancer. The activecompounds may preferably be selected from the group comprising radiationtherapeutics, chemotherapeutics including but not limited totemozolomide, vincristine, vinorelbine, procarbazine, carmustine,lomustine, taxol, peloruside, taxotere, tamoxifen, retinoic acid,5-fluorouracil, cyclophosphamide and thalidomide.

The compound or the pharmaceutical composition as defined herein canthus be administered alone or in combination with one or more activecompounds. The latter can be administered before, after orsimultaneously with the administration of the said agent(s).

The term “pharmaceutically acceptable” as used herein is consistent withthe art and means compatible with the other ingredients of apharmaceutical composition and not deleterious to the recipient thereof.

The compounds as defined herein can optionally be formulated into apharmaceutically acceptable salt. The term “pharmaceutically acceptablesalt” as used herein means an inorganic acid addition salt such ashydrochloride, sulfate, and phosphate, or an organic acid addition saltsuch as acetate, maleate, fumarate, tartrate, and citrate. Examples ofpharmaceutically acceptable metal salts are alkali metal salts such assodium salt and potassium salt, alkaline earth metal salts such asmagnesium salt and calcium salt, aluminum salt, and zinc salt. Examplesof pharmaceutically acceptable ammonium salts are ammonium salt andtetramethylammonium salt. Examples of pharmaceutically acceptableorganic amine addition salts are salts with morpholine and piperidine.Examples of pharmaceutically acceptable amino acid addition salts aresalts with lysine, glycine, and phenylalanine.

The pharmaceutical composition as defined herein may further comprise atleast one active compound, as defined above.

The pharmaceutical composition as defined herein can be administeredorally, for example in the form of pills, tablets, lacquered tablets,sugar-coated tablets, granules, hard and soft gelatin capsules, aqueous,alcoholic or oily solutions, syrups, emulsions or suspensions, orrectally, for example in the form of suppositories. Administration canalso be carried out parenterally, for example subcutaneously,intramuscularly or intravenously in the form of solutions for injectionor infusion. Other suitable administration forms are, for example,percutaneous or topical administration, for example in the form ofointments, tinctures, sprays or transdermal therapeutic systems, or theinhalative administration in the form of nasal sprays or aerosolmixtures, or, for example, microcapsules, implants or rods.

The pharmaceutical composition as defined herein can be prepared in amanner known per se to one of skill in the art. For this purpose, atleast one compound as defined herein or a pharmaceutically acceptablesalt thereof as defined above, one or more solid or liquidpharmaceutical excipients and, if desired, in combination with otherpharmaceutical active compounds, can be brought into a suitableadministration form or dosage form which can then be used as apharmaceutical in human medicine or veterinary medicine.

By means of non-limiting examples, such a formulation may be in a formsuitable for oral administration, for parenteral administration (such asby intravenous, intramuscular, or subcutaneous injection, or intravenousinfusion), for topical administration (including ocular), foradministration by inhalation, by a skin patch, by an implant, by asuppository, etc. Such suitable administration forms—which may be solid,semi-solid, or liquid, depending on the manner of administration—as wellas methods and carriers, diluents and excipients for use in thepreparation thereof, will be clear to the skilled person as e.g.disclosed in standard textbooks, such as the latest edition ofRemington's Pharmaceutical Sciences.

As non-limiting examples, the active compound as defined herein,together with one or more solid or liquid pharmaceutical carriersubstances and/or additives (or auxiliary substances) and, if desired,in combination with other pharmaceutically active compounds havingtherapeutic or prophylactic action, can be brought into a suitableadministration form or dosage form which can then be used as apharmaceutical in human medicine. For the production of pills, tablets,sugar-coated tablets and hard gelatin capsules it is possible to use,for example, lactose, starch, for example maize starch, or starchderivatives, talc, stearic acid or its salts, etc. Carriers for softgelatin capsules and suppositories are, for example, fats, waxes,semisolid and liquid polyols, natural or hardened oils, etc. Suitablecarriers for the preparation of solutions, for example of solutions forinjection, or of emulsions or syrups are, for example, water,physiological sodium chloride solution, alcohols such as ethanol,glycerol, polyols, sucrose, invert sugar, glucose, mannitol, vegetableoils, etc. It is also possible to lyophilize the nucleic acid and/or theactive compound and to use the resulting lyophilisates, for example, forpreparing preparations for injection or infusion. Suitable carriers formicrocapsules, implants or rods are, for example, copolymers of glycolicacid and lactic acid.

The pharmaceutical compositions as defined herein can also containadditives, for example fillers, disintegrants, binders, lubricants,wetting agents, stabilizers, emulsifiers, dispersants, preservatives,sweeteners, colorants, flavorings, aromatizers, thickeners, diluents,buffer substances, solvents, solubilizers, agents for achieving a depoteffect, salts for altering the osmotic pressure, coating agents orantioxidants.

For an oral administration form, the pharmaceutical compositions asdefined herein can be mixed with suitable additives, such as excipients,stabilizers, or inert diluents, and brought by means of the customarymethods into the suitable administration forms, such as tablets, coatedtablets, hard capsules, aqueous, alcoholic, or oily solutions. Examplesof suitable inert carriers are gum arabic, magnesia, magnesiumcarbonate, potassium phosphate, lactose, glucose, or starch, inparticular, corn starch. In this case, the preparation can be carriedout both as dry and as moist granules. Suitable oily excipients orsolvents are vegetable or animal oils, such as sunflower oil or codliver oil. Suitable solvents for aqueous or alcoholic solutions arewater, ethanol, sugar solutions, or mixtures thereof. Polyethyleneglycols and polypropylene glycols are also useful as further auxiliariesfor other administration forms. As immediate release tablets, thesecompositions may contain microcrystalline cellulose, dicalciumphosphate, starch, magnesium stearate, and lactose and/or otherexcipients, binders, extenders, disintegrants, diluents, and lubricantsknown in the art.

The oral administration of a pharmaceutical composition as definedherein comprising at least one compound as defined herein, or apharmaceutically acceptable salt or ester and/or solvate thereof, issuitably accomplished by uniformly and intimately blending together asuitable amount of said compound in the form of a powder, optionallyalso including a finely divided solid carrier, and encapsulating theblend in, for example, a hard gelatin capsule. The solid carrier caninclude one or more substances, which act as binders, lubricants,disintegrating agents, coloring agents, and the like. Suitable solidcarriers include, for example, calcium phosphate, magnesium stearate,talc, sugars, lactose, dextrin, starch, gelatin, cellulose,polyvinylpyrrolidine, low melting waxes and ion exchange resins.

Some preferred, but non-limiting examples of such preparations includetablets, pills, powders, lozenges, sachets, cachets, elixirs,suspensions, emulsions, solutions, syrups, aerosols, ointments, cremes,lotions, soft and hard gelatin capsules, suppositories, drops, sterileinjectable solutions and sterile packaged powders (which are usuallyreconstituted prior to use) for administration as a bolus and/or forcontinuous administration, which may be formulated with carriers,excipients, and diluents that are suitable per se for such formulations,such as lactose, dextrose, sucrose, sorbitol, mannitol, starches, gumacacia, calcium phosphate, alginates, tragacanth, gelatin, calciumsilicate, microcrystalline cellulose, polyvinylpyrrolidone, polyethyleneglycol, cellulose, (sterile) water, methylcellulose, methyl- andpropylhydroxybenzoates, talc, magnesium stearate, edible oils, vegetableoils and mineral oils or suitable mixtures thereof. The formulations canoptionally contain other pharmaceutically active substances (which mayor may not lead to a synergistic effect with the compounds of theinvention) and other substances that are commonly used in pharmaceuticalformulations, such as lubricating agents, wetting agents, emulsifying,and suspending agents, dispersing agents, desintegrants, bulking agents,fillers, preserving agents, sweetening agents, flavoring agents, flowregulators, release agents, etc. The compositions may also be formulatedso as to provide rapid, sustained, or delayed release of the activecompound(s) contained therein, for example using liposomes orhydrophilic polymeric matrices based on natural gels or syntheticpolymers.

The dosage or amount of compounds of the invention used, optionally incombination with one or more active compounds to be administered,depends on the individual case and is, as is customary, to be adapted tothe individual circumstances to achieve an optimum effect. Thus, itdepends on the nature and the severity of the disorder to be treated,and also on the sex, age, weight and individual responsiveness of thehuman or animal to be treated, on the efficacy and duration of action ofthe compounds used, on whether the therapy is acute or chronic orprophylactic, or on whether other active compounds are administered inaddition to the agent(s) of the invention.

In another embodiment, the invention provides a kit comprising apharmaceutical composition as defined herein, and a further activecompound as defined herein, for simultaneous, separate or sequentialadministration to a subject in need thereof.

In accordance with the method of treatment as defined herein, saidpharmaceutical composition can be administered separately at differenttimes during the course of therapy or concurrently in divided or singlecombination forms. The treatment method is therefore to be understood asembracing all such regimes of simultaneous or alternating treatment andthe term “administering” is to be interpreted accordingly.

Essentially, the primary modes of treatment of solid tumor cancerscomprise surgery, radiation therapy, and chemotherapy, separately and incombination. The compounds as defined herein are suitable for use incombination with these medicinal techniques. The compounds as definedherein may be useful in increasing the sensitivity of tumor cells toradiation in radiotherapy and also in potentiating or enhancing damageto tumors by chemotherapeutic agents. The compounds as defined hereinand their pharmaceutically acceptable salts and/or solvates may also beuseful for sensitizing multidrug-resistant tumor cells. The compounds asdefined herein are useful therapeutic compounds for administration inconjunction with other drugs or therapies to potentiate their effect.

In another embodiment of the method of the invention, the administrationmay be performed with food, e.g., a high-fat meal. The term “with food”means the consumption of a meal either during or no more than about onehour before or after administration of a pharmaceutical compositionaccording to the invention.

Oral administration of a pharmaceutical composition comprising at leastone compound as defined herein, or a pharmaceutically acceptable salt orester and/or solvate thereof can also be accomplished by preparingcapsules or tablets containing the desired amount of said compound,optionally blended with a solid carrier as described above. Compressedtablets containing the pharmaceutical composition of the invention canbe prepared by uniformly and intimately mixing the active ingredientwith a solid carrier such as described above to provide a mixture havingthe necessary compression properties, and then compacting the mixture ina suitable machine to the shape and size desired. Molded tablets may bemade by molding in a suitable machine, a mixture of powdered compoundmoistened with an inert liquid diluent.

When administered by nasal aerosol or inhalation, these compositions maybe prepared according to techniques well-known in the art ofpharmaceutical formulation and may be prepared as solutions in saline,employing benzyl alcohol or other suitable preservatives, absorptionpromoters to enhance bioavailability, fluorocarbons, and/or othersolubilizing or dispersing agents known in the art. Suitablepharmaceutical formulations for administration in the form of aerosolsor sprays are, for example, solutions, suspensions, or emulsions of thecompounds of the invention or their physiologically tolerable salts in apharmaceutically acceptable solvent, such as ethanol or water, or amixture of such solvents. If required, the formulation can alsoadditionally contain other pharmaceutical auxiliaries such assurfactants, emulsifiers and stabilizers as well as a propellant.

For subcutaneous or intravenous administration, the compound as definedherein, if desired with the substances customary therefore such assolubilizers, emulsifiers, or further auxiliaries, are brought intosolution, suspension, or emulsion. The compounds as defined herein canalso be lyophilized and the lyophilizates obtained used, for example,for the production of injection or infusion preparations. Suitablesolvents are, for example, water, physiological saline solution, oralcohols, e.g. ethanol, propanol, glycerol, in addition also sugarsolutions such as glucose or mannitol solutions, or alternativelymixtures of the various solvents mentioned. The injectable solutions orsuspensions may be formulated according to known art, using suitablenon-toxic, parenterally-acceptable diluents, or solvents, such asmannitol, 1,3-butanediol, water, Ringer's solution, or isotonic sodiumchloride solution, or suitable dispersing or wetting and suspendingagents, such as sterile, bland, fixed oils, including synthetic mono- ordiglycerides, and fatty acids, including oleic acid.

When rectally administered in the form of suppositories, theseformulations may be prepared by mixing the compounds according to theinvention with a suitable non-irritating excipient, such as cocoabutter, synthetic glyceride esters, or polyethylene glycols, which aresolid at ordinary temperatures, but liquidify and/or dissolve in therectal cavity to release the drug.

The pharmaceutical compositions as defined herein can be administered tohumans in dosage ranges specific for each compound comprised in saidcompositions. The compounds comprised in said composition can beadministered together or separately.

It will be understood, however, that specific dose level and frequencyof dosage for any particular patient may be varied and will depend upona variety of factors including the activity of the specific compoundemployed, the metabolic stability and length of action of that compound,the age, body weight, general health, sex, diet, mode and time ofadministration, rate of excretion, drug combination, the severity of theparticular condition, and the host undergoing therapy.

The present invention also provides a new synthetic route to obtain thecompounds as defined herein in large quantities.

Said new process for producing a compound according to general formula(I) of the invention, comprises the steps of:

-   -   (a) reacting a methyl-ketone having structural Formula IIa with        an aldehyde having structural Formula IIb, through aldol        coupling, wherein P⁵ is a protecting group, executing suitable        functional group interconversions, thereby obtaining a compound        having structural Formula II;    -   (b) protecting the functional groups at positions X³ and X⁴ in        the compound of Formula II where needed;    -   (c) reacting the compound of Formula II with a compound of the        Formula III in the presence of a suitable catalyst, giving rise        to a compound of the formula (IV),    -   (d) removing the protecting groups P¹ and P⁴ in the resulting        compound and esterifying the deprotected COOH with the C₁₅ OH        group, executing suitable functional group interconversions,        thereby obtaining the lactone having structural Formula V; and    -   (e) deprotecting P⁵ in the compound having structural Formula V        and deprotecting any of the possibly protected X¹, X², X³, or X⁴        groups, if required after executing additional suitable        functional group interconversions, thereby obtaining a compound        having structural Formula I, generally following the reaction        scheme below:

-   -   wherein    -   P¹ is hydrogen or a carboxyl protecting group; for example        optionally substituted alkyl, preferably P¹ is methyl;    -   P² is selected from halogen, pseudohalogen, CF₃SO₃, OAc,        preferably P² is bromo, and simultaneously P³ is a trialkyltin,        such as trimethyltin or tri-n-butyltin, a boronic acid or a        boronic ester;    -   or, alternatively,    -   P² is a trialkyltin, such as trimethyltin or tri-n-butyltin, a        boronic acid or ester, and simultaneously P³ is selected from        halogen, pseudohalogen, CF₃SO₃, OAc, (for example P³ is bromo);    -   P⁴ is selected from an orthogonally chosen protecting group,        preferably P⁴ is MPM (4-OMe-Bn);    -   P⁵ is a protecting group which can be orthogonally removed,        preferably selected from: TBS (tert-butyldimethylsilyl) or MEM        (2-methoxyethoxymethyl);    -   the suitable catalyst is typically a transition metal with        ligands. For example, said suitable catalyst can be        Pd₂(dba)₃.CHCl₃; and X¹, X², X³, and X⁴ have the same meaning as        that defined above, and wherein, if present, their functional        groups are suitably protected in steps (a) and (b), functionally        interconverted where needed and deprotected accordingly in step        (e).

Depending on the type of functional group or substituent on X¹, X², X³,and X⁴, there may be a different need to shield them from reacting withthe other functional groups during the different coupling steps or toselectively prevent them to undergo a functional group interconversion.It may hence be needed to e.g. protect the functional groups of X¹, X²,X³, and X⁴ in steps (a) and (b) and subsequently deprotect themaccordingly in final step (e).

Such protective groups are known in the art. Preferred but non-limitingprotective groups can be selected from the group comprising:

a) for hydroxyl groups: Methoxymethyl ether (MOM-OR),(2-Methoxyethoxy)methyl ether (MEM-OR), Tetrahydropyranyl ether(THP-OR), t-Butyl ether, Allyl ether, Benzyl ether (Bn-OR),t-Butyldimethylsilyl ether (TBDMS-OR), t-Butyldiphenylsilyl ether(TBDPS-OR), Acetic acid ester, Pivalic acid ester, Benzoic acid ester,and the like;b) for carbonyl groups: Dimethyl acetal, 1,3-Dioxane, 1,3-Dioxolane,1,3-Dithiane, N,N-Dimethylhydrazone, and the like;c) for carboxyl groups: Methyl ester, Ethyl ester, t-Butyl ester, Benzylester, S-t-Butyl ester, 2-Alkyl-1,3-oxazoline, and the like;d) for amino groups: 9-Fluorenylmethyl carbamate (Fmoc-NRR′), t-Butylcarbamate (Boc-NRR′), Benzyl carbamate (Z—NRR′, Cbz-NRR′), Acetamide,Trifluoroacetamide, Phthalimide, Benzylamine (Bn-NRR′),Triphenylmethylamine (Tr-NRR′), Benzylideneamine, p-Toluenesulfonamide(Ts-NRR′), and the like.

The present invention is further illustrated by the following examples,which do not limit the scope of the invention in any way.

EXAMPLES Example 1 First Synthesis of JC168

The first synthesis of JC168 was based on the aldol reaction between anenantiomerically pure aldehyde with an enantiomerically puremethylketone. This aldol reaction resulted in the wrong stereochemistryat C₁₃ and C₁₅. Therefore, extra steps (an oxidation/reduction sequenceat C₁₃ and a Mitsunobu inversion at C₁₅) were needed to incorporate thecorrect stereochemistry. Other key steps involve two consecutive Stillecouplings, asymmetric dihydroxylation and macrolactonization.

(1′Z,3R,3′R,4S,7S,9S,11S)-3,4,11-Trihydroxy-7-(3′-hydroxymethyl-1′-methylpent-1′-enyl)-9-methoxy-12,12-dimethyl-6-oxabicyclo[11.3.1]heptadeca-1(17),13,15-trien-5-one

Formula: C₂₆H₄₀O₇

Molar Mass: 464.6 g/mol

R_(f): 0.26 (pentane/acetone 6/4)

The ¹H NMR spectrum of JC168 is provided in FIG. 6.

¹H NMR (300 MHz, C₆D₆): δ 7.31 (1H, s), 7.07 (2H, s with finestructure), 6.96-6.90 (1H, m), 5.61 (1H, dd, 3.4 Hz and 10.4 Hz), 4.81(1H, d 10.4 Hz), 4.40-4.36 (1H, m), 3.65 (1H, 3.8 Hz and 10.6 Hz),3.60-3.55 (1H, m), 3.15-3.09 (4H, m), 3.09-2.73 (1H, m), 2.73-2.61 (1H,m), 2.53-2.42 (1H, m), 1.82-1.55 (6H, m), 1.36-1.12 (10H, m), 1.01-0.86(2H, m), 0.75 (3H, t, 7.3 Hz)

The ¹³C NMR spectrum of JC168 is provided in FIG. 7.

¹³C NMR (75 MHz, C₆D₆): δ 174.48 (C), 147.14 (C), 137.69 (C), 137.35(C), 130.40 (CH), 129.22 (CH), 128.86 (CH), 128.39-127.74 (C6D6+CH),124.69 (CH), 76.74 (CH), 75.23 (CH), 74.28 (CH), 73.94 (CH), 69.52 (CH),67.25 (CH2), 56.97 (CH3), 43.77 (CH), 42.60 (CH2), 41.21 (CH2), 38.91(CH2), 35.45 (CH2), 26.42 (CH3), 24.90 (CH2), 21.63 (CH3), 18.30 (CH3),12.38 (CH3)

ESI-MS (m/z): 465.2 (M+H+)

Example 2 New Synthesis of JC168

The general synthesis route was described above and is now exemplifiedfor compound JC 168. The process starts with the coupling of 2fragments: a methylketone that already possesses the correctstereochemistry at C₁₅ and C₁₈, and a prochiral aldehyde (cf. Schemebelow). Aldol coupling of those 2 fragments delivers a hydroxyketonewith the correct stereochemistry at C₁₁. Key steps involve asamarium-mediated anti-reduction of this hydroxyketone, securing thecorrect stereochemistry at C₁₃, consecutive Stille couplings to completethe carbon-skeleton of pelofen, asymmetric dihydroxylation andmacrolactonization.

Compound J27

(3′S,5′R,7′S,10′R,Z)-5′-methoxy-7′-((4″-methoxybenzyl)oxy)-10′-(((2′″-methoxyethoxy)methoxy)methyl)-2′,8′-dimethyl-2′-(3″″-(trimethylstannyl)phenyl)dodec-8′-en-3′-ylpropionate

Formula: C₄₀H₆₄O₈Sn

Molar Mass: 791.65 g/mol

R_(f): 0.29 (pentane/diethylether 6/4)

¹H NMR (500 MHz, CDCl₃) δ 7.48 (app. d, J=2.2 Hz, 1H), 7.35-7.23 (m,3H), 7.16 (app. d, J=8.7 Hz, 2H), 6.84 (app. d, J=8.7 Hz, 2H), 5.51 (dd,J=9.8, 2.1 Hz, 1H), 5.12 (dd, J=10.3, 1.4 Hz, 1H), 4.69 (d, J=6.7 Hz,1H, A part of AB-spinsystem), 4.68 (d, J=6.7 Hz, 1H, B part ofAB-spinsystem), 4.31 (d, J=11.1 Hz, 1H), 4.20 (dd, J=9.9, 2.9 Hz, 1H),4.04 (d, J=11.2 Hz, 1H), 3.80 (s, 3H), 3.68-3.65 (m, 2H), 3.56-3.53 (m,2H), 3.46 (dd, J=9.5, 5.7 Hz, 1H), 3.39 (dd, J=9.6, 6.7 Hz, 1H), 3.37(s, 3H), 3.22 (s, 3H), 3.18-3.13 (m, 1H), 2.51-2.42 (m, 1H), 2.22 (app.q, J=7.6 Hz, 2H), 2.03 (ddd, J=14.1, 10.0, 4.0 Hz, 1H), 1.66 (d, J=1.4Hz, 3H), 1.52-1.40 (m, 3H), 1.32-1.26 (m, 1H), 1.31 (s, 3H), 1.29 (s,3H), 1.19-1.09 (m, 1H), 1.04 (app. t, J=7.6 Hz, 3H), 0.76 (app. t, J=7.5Hz, 3H), 0.27 (s, 9H)

¹³C NMR (125 MHz, CDCl₃) δ 174.00 (Cq), 159.12 (Cq), 145.86 (Cq), 141.76(Cq), 136.44 (Cq), 133.84 (CH), 133.71 (CH), 131.23 (Cq), 131.12 (CH),129.19 (2×CH), 127.81 (CH), 126.72 (CH), 113.84 (2×CH), 95.66 (CH₂),76.79 (CH), 75.05 (CH), 73.36 (CH), 71.93 (CH₂), 71.51 (CH₂), 69.70(CH₂), 66.87 (CH₂), 59.18 (CH₃), 56.46 (CH₃), 55.42 (CH₃), 41.95 (Cq),39.26 (CH), 37.67 (CH₂), 35.18 (CH₂), 27.93 (CH₂), 26.03 (CH₃), 25.21(CH₂), 23.66 (CH₃), 18.04 (CH₃), 11.91 (CH₃), 9.52 (CH₃), −9.41 (3×CH₃)

Compound J28

methyl(E)-4-(3′-((3″S,5″R,7″S,10″R,Z)-5″-methoxy-7″-((4′″-methoxybenzyl)oxy)-10″-(((2″″-methoxyethoxy)methoxy)methyl)-2″,8″-dimethyl-3″-(propionyloxy)dodec-8″-en-2″-yl)phenyl)but-2-enoate

Formula: C₄₂H₆₂O₁₀

Molar Mass: 726.94 g/mol

R_(f): 0.31 (pentane/diethylether 4/6)

¹H NMR (500 MHz, CDCl₃) δ 7.26-7.15 (m, 5H), 7.08 (app. dt, J=15.6, 6.8Hz, 1H), 6.98 (app. dt, J=6.9, 1.7 Hz, 1H), 6.85 (app. d, J=8.7 Hz, 2H),5.80 (app. dt, J=15.6, 1.6 Hz, 1H), 5.47 (dd, J=10.1, 1.9 Hz, 1H), 5.12(dd, J=10.4, 1.5 Hz, 1H), 4.68 (d, J=6.7 Hz, 1H, A part ofAB-spinsystem), 4.67 (d, J=6.7 Hz, 1H, B part of AB-spinsystem), 4.32(d, J=11.3 Hz, 1H), 4.20 (dd, J=10.0, 3.0 Hz, 1H), 4.05 (d, J=11.3 Hz,1H), 3.80 (s, 3H), 3.70 (s, 3H), 3.68-3.65 (m, 2H), 3.56-3.53 (m, 2H),3.48 (app. dd, J=6.7, 1.8 Hz, 2H), 3.46 (dd, J=9.2, 5.9 Hz, 1H), 3.39(dd, J=9.4, 6.7 Hz, 1H), 3.37 (s, 3H), 3.21 (s, 3H), 3.18-3.12 (m, 1H),2.51-2.42 (m, 1H), 2.21 (app. q, J=7.6 Hz, 2H), 2.03 (ddd, J=14.2, 10.2,3.9 Hz, 1H), 1.67 (d, J=1.4 Hz, 3H), 1.52-1.38 (m, 3H), 1.33-1.26 (m,1H), 1.29 (s, 3H), 1.27 (s, 3H), 1.19-1.09 (m, 1H), 1.04 (app. t, J=7.6Hz, 3H), 0.77 (app. t, J=7.4 Hz, 3H)

¹³C NMR (125 MHz, CDCl₃) δ 173.94 (Cq), 167.13 (Cq), 159.12 (Cq), 147.92(CH), 147.07 (Cq), 137.33 (Cq), 136.46 (Cq), 131.23 Cq), 131.11 (CH),129.15 (2×CH), 128.50 (CH), 127.20 (CH), 126.63 (CH), 125.14 (CH),121.95 (CH), 113.84 (2×CH), 95.66 (CH₂), 76.65 (CH), 74.97 (CH), 73.29(CH), 71.93 (CH₂), 71.50 (CH₂), 69.65 (CH₂), 66.88 (CH₂), 59.18 (CH₃),56.44 (CH₃), 55.42 (CH₃), 51.57 (CH₃), 41.87 (Cq), 39.28 (CH), 38.87(CH₂), 37.61 (CH₂), 35.12 (CH₂), 27.91 (CH₂), 25.74 (CH₃), 25.22 (CH₂),23.84 (CH₃), 18.03 (CH₃), 11.90 (CH₃), 9.49 (CH₃)

Example 3 Synthesis of C₂-C₃ analogue J74 3.1 Synthesis of J20Preparation of (−)-Diisopinocampheylmethoxyborane:

To a stirring solution of (+)-α-pinene, freshly distilled over CaH₂,(17.7 ml, 112 mmol, 2.5 eq.) in THF (13.4 ml) was added BH₃.SMe₂ (4.2ml, 45 mmol, 1 eq.) over 30 minutes at a constant temperature of 23° C.After the addition, the stirring was stopped and the solution was leftuntouched overnight, during which white crystals were formed. Thesolution was then cooled to 0° C. for 2 h, after which the supernatantswas removed using a double-ended needle. The white crystals were washedwith precooled, dry Et₂O (3×8 ml) at 0° C., and dried, first by blowingAr, then overnight in vacuo, yielding (−)-diisopinocampheylborane (11.4g, 40 mmol, 89%).

(−)-diiospinocampheylborane (11.4 g, 40 mmol, 1 eq.), was suspended inEt₂O (40 ml) and cooled to 0° C. MeOH (1.6 ml, 40 mmol, 1 eq.) was thenadded dropwise and the solution was stirred at RT until everything haddissolved.

Allylation:

The solution of (−)-diisopinocampheylmethoxyborane (12.6 g, 40 mmol, 2.4eq) in Et₂O (40 ml) was diluted with Et₂O (143 ml) and cooled to −78° C.Allylmagnesiumbromide (40 ml, 40 mmol, 1M in Et₂O) was added dropwise,upon which the reaction mixture became turbid and a black aggregate wasformed. The reaction mixture was first stirred for 15 min at −78° C.,followed by 1 h of stirring at RT, resulting in a white solution. Thereaction mixture was then cooled back to −78° C., and a solution of thestarting material (3.8 g, 17 mmol, 1 eq.) in Et₂O (26 ml) at −78° C. wasadded via a double-ended needle. The reaction mixture was stirredovernight during which the temperature was allowed to rise from −78° C.to −30° C. Tlc-analysis after 13 h (CH₂Cl₂/acetone 9/1) showed completeconversion of the starting material. The reaction was quenched by addinga solution of NaOOH in water. This solution was prepared beforehand byadding H₂O₂ (10.5 ml, 119 mmol, 7.2 eq.) to a solution of NaOH (3.2 g,80 mmol, 4.8 eq.) in H₂O (26 ml) at 0° C. in a separate flask. TheNaOH-solution was transferred to the reaction mixture at 0° C., afterwhich it was stirred at RT for 4 h. The reaction mixture was then pouredin aq. NH₄Cl (100 ml), the phases were separated and the aqueous phasewas extracted with EtOAc (3×200 ml). The combined organic phases weredried over MgSO₄ and concentrated. Purification was accomplished byconsecutive flash column chromatography (CH₂Cl₂/acetone 95/5;Hexane/acetone 8/2; CH₂Cl₂/acetone 88/12), yielding the target materialJ20 (3.6 g, 79%) as a clear, colorless oil.

Z-(4S,7R)-7-(((2′-methoxyethoxy)methoxy)methyl)-5-methylnona-1,5-diene-4-ol

Formula: C₁₅H₂₈O₄

Molar mass: 272.38

R_(f): 0.15 (CH₂Cl₂/acetone 9/1)

ESI MS: 255.2 (M−H₂O+H⁺)

[α]_(D): −6.6° (c=8.4 mg/ml in CHCl₃)

IR (HATR): 3435 (b), 2929, 2876, 2186 (w), 2009 (w), 1982 (w), 1451,1408, 1376, 1292, 1242, 1199, 1174, 1111 (m), 1095, 1044 (s), 1019 (s),989 (m), 938, 911, 852 cm-1

¹H NMR (300 MHz, C6D6) δ 5.91 (app. dt, J=17.2, 10.3, 6.9 Hz, 1H), 5.10(app. ddt, J=17.2, 3.6, 1.8 Hz, 1H), 5.03 (app. ddt, J=10.2, 2.3, 1.1Hz, 1H), 4.94 (app. dd, J=9.9, 1.8 Hz, 1H), 4.64 (app. t, J=7.2 Hz, 1H),4.48 (app. s, 2H), 3.59-3.46 (m, 2H), 3.36 (dd, J=9.1, 4.9 Hz, 1H),3.33-3.26 (m, 2H), 3.14 (dd, J=17.9, 9.0 Hz, 1H), 3.11 (s, 3H),2.65-2.50 (m, 1H), 2.39-2.28 (m, 1H), 1.89 (d, J=1.5 Hz, 3H), 1.36-1.21(m, 1H), 1.14-0.97 (m, 1H), 0.78 (app. t, J=7.3 Hz, 3H)

¹³C NMR (75 MHz, C6D₆) δ 140.46 (Cq), 136.31 (CH), 131.13 (CH), 116.87(CH2), 95.86 (CH2), 72.55 (CH2), 71.76 (CH2), 69.25 (CH), 67.62 (CH2),58.97 (CH3), 39.84 (CH), 39.37 (CH2), 25.60 (CH2), 18.60 (CH3), 12.29(CH3)

3.2 Synthesis of J23

To a flask containing NaH (1.05 g, 26 mmol, 2 eq., 60 m % dispersion inmineral oil) was added a solution of the starting material J20 (3.58 g,13 mmol, 1 eq.) in THF (40 ml) at 0° C. The reaction mixture was stirredfor 30 min at RT, after which methoxyphenylmethyl-chloride (3.6 ml, 26mmol, 2 eq.) was added dropwise via syringe, followed by a suspension oftetrabutylammoniumiodide (9.7 g, 26 mmol, 2 eq) in DMF (13 ml). Thereaction was stirred at RT for 5 h, after which tlc-analysis(CH₂Cl₂/acetone 9/1) showed complete conversion of the startingmaterial. The reaction mixture was then gently poured in a separationfunnel containing water (200 ml), and extracted with Et₂O (4×200 ml).The combined organic fractions were dried over MgSO₄, and concentrated.The product was purified by flash column chromatography (pentane/Et₂O8/2), yielding the desired product J23 as a clear yellow oil (5.10 g, 13mmol, 99%)

Z-(4S,7R)-4-((4″-methoxybenzyl)oxy)-7-(((2′-methoxyethoxy) methoxy)methyl)-5-methylnona-1,5-diene

Formula: C₂₃H₃₆O₅

Molar mass: 392.54

R_(f): 0.31 (pentane/Et₂O 6/4)

ESI MS: 410.2 (M+NH₄ ⁺)

[α]_(D): −66.9° (c=7.6 mg/ml in CHCl₃)

IR (HATR): 3008 (w), 2933 (w), 2876 (w), 1613 (w), 1513, 1462 (w), 1455(w), 1300 (w), 1247, 1214, 1172, 1110, 1095, 1075, 1049 (m), 991 (w),916 (w), 847 (w), 823 (w), 751 (s), 666 cm⁻¹

¹H NMR (300 MHz, C6D6) δ 7.39 (app. d, J=8.6 Hz, 2H), 6.86 (app. d,J=8.9 Hz, 2H), 5.96 (dddd, J=17.1, 10.2, 7.4, 6.8 Hz, 1H), 5.20 (dd,J=10.3, 2.0 Hz, 1H), 5.10 (app. ddd, J=17.1, 3.6, 1.4 Hz, 1H), 5.04(app. ddt, J=10.1, 2.2, 1.1 Hz, 1H), 4.61 (app. s, 2H), 4.60 (d, J=11.7Hz, 1H), 3.39 (dd, J=8.1, 5.8 Hz, 1H), 4.36 (d, J=11.7 Hz, 1H),3.64-3.60 (m, 2H), 3.42 (app. dd, J=6.4, 3.1 Hz, 2H), 3.39-3.34 (m, 2H),3.32 (s, 3H), 3.12 (s, 3H), 2.71-2.52 (m, 2H), 3.42 (app. dddt, J=14.1,7.3, 5.9, 1.3 Hz, 1H), 1.80 (d, J=1.3 Hz, 3H), 1.61-1.46 (m, 1H),1.27-1.11 (m, 1H), 0.83 (app. t, J=7.4 Hz, 3H)

¹³C NMR (75 MHz, C6D6) δ 160.98 (Cq), 136.84 (Cq), 136.34 (CH), 132.35(CH+Cq), 129.67 (2×CH), 116.81 (CH2), 114.42 (2×CH), 96.12 (CH2), 77.12(CH), 72.61 (CH2), 72.07 (CH2), 70.18 (CH2), 67.59 (CH2), 59.03 (CH3),55.12 (CH3), 39.79 (CH), 39.54 (CH2), 25.92 (CH2), 18.47 (CH3), 12.33(CH3)

3.3 Synthesis of J3

To a solution of the diene J23 (5.10 g, 13 mmol, 1 eq.) in a mixture ofDMF (240 ml) and water (24 ml) was added Cu(OAc)₂ (3.94 g, 20 mmol, 1.5eq.) in one portion and PdCl₂ (582 mg, 3.3 mmol, 0.25 eq.) in differentportions. The reaction mixture was bubbled using O₂ and stirred at roomtemperature until tlc-analysis showed complete conversion of thestarting material. The reaction mixture was then poured in water (500ml) and extracted with Et₂O (3×800 ml). The combined organic phases weredried over MgSO₄ and concentrated. After concentration, the crudeproduct was filtered over a patch of Celite®, rinsed with Et₂O (200 ml),and transferred to a separation funnel containing water (300 ml) usingEt₂O (100 ml). The phases were separated and the aqueous phase wasextracted with Et₂O (2×300 ml). Again, the combined organic phases weredried over MgSO₄ and concentrated. Purification via flash columnchromatography (pentane/Et₂O 6/4) delivered methyl ketone J3 as a clearyellow oil (4.88 g, 12 mmol, 91%).

Z-(4S,7R)-4-((4″-methoxybenzyl)oxy)-7-(((2′-methoxyethoxy)methoxy)methyl)-5-methylnon-5-en-2-one

Formula: C₂₃H₃₆O₆

Molar mass: 408.53

R_(f): 0.16 (pentane/diethylether 6/4)

ESI MS: 426.2 (M+NH₄); 431.2 (M+Nat)

[α]_(D): −43.8° (c=8.0 mg/ml in CHCl₃)

IR (HATR): 3013 (w), 2957 (w), 2933 (w), 2871 (w), 1714, 1611 (w), 1514,1462 (w), 1454 (w), 1358 (w), 1300 (w), 1248, 1216, 1173, 1092, 1049,850 (w), 822 (w), 752 (s), 666 (w) cm⁻¹

¹H NMR (300 MHz, C6D6) δ 7.33 (app. d, J=8.3 Hz, 2H), 6.82 (app. d,J=8.4 Hz, 2H), 5.18 (app. d, J=10.2 Hz, 1H), 5.01 (dd, J=9.4, 2.9 Hz,1H), 4.60 (app. s, 2H), 4.55 (d, J=11.4 Hz, 1H), 4.38 (d, J=11.4 Hz,1H), 3.66-3.58 (m, 2H), 3.48-3.34 (m, 4H), 3.30 (s, 3H), 3.13 (s, 3H),2.78 (dd, J=15.4, 9.4 Hz, 1H), 2.78-2.65 (m, 1H), 2.12 (dd, J=15.4, 3.0Hz, 1H), 1.80 (s, 3H), 1.75 (d, J=1.0 Hz, 3H), 1.59-1.43 (m, 1H),1.24-1.07 (m, 1H), 0.81 (app. t, J=7.4 Hz, 3H)

¹³C NMR (75 MHz, C6D6) δ 205.02 (Cq), 160.04 (Cq), 136.12 (Cq), 132.60(CH), 131.90 (Cq), 129.89 (2×CH), 114.42 (2×CH), 96.11 (CH2), 73.90(CH), 72.62 (CH2), 72.02 (CH2), 70.60 (CH2), 67.60 (CH2), 59.03 (CH3),55.11 (CH3), 48.30 (CH2), 39.98 (CH), 31.17 (CH3), 25.76 (CH2), 18.54(CH3), 12.27 (CH3)

3.4 Synthesis of J4

To a stirred solution of ketone J3 (353 mg, 0.86 mmol, 1 eq.) in Et₂O(10 ml) was added triethylamine (205 μl, 1.47 mmol, 1.7 eq.) at RT. Thesolution was then cooled to 0° C. and chlorodicylcohexylborane (1.3 ml,1.3 mmol, 1.5 eq., 1 M in hexane) was added dropwise, upon which thereaction mixture became turbid. After stirring for 30 min at 0° C., thereaction mixture was further cooled to −15° C. A solution of aldehyde(588 mg, 2.59 mmol, 3 eq.) in Et₂O (2 ml) was then added via adouble-ended needle and the flask containing the aldehyde was rinsedwith Et₂O (3×1 ml). The reaction mixture was stirred for 20 h at −11°C., after which tlc-analysis showed complete conversion of the startingmaterial (hexane/acetone 8/2). The reaction was quenched by consecutiveaddition of pH 7 phosphate buffer (9 ml), MeOH (9 ml) and H₂O₂ (1 ml, 13mmol, 15 eq., 35 v % in H₂O) and stirred for 3 h at room temperature.The reaction mixture was then poured into a separation funnel containingwater (100 ml), extracted with CH₂Cl₂ (3×100 ml), dried over MgSO₄ andconcentrated. Purification using gradient flash column chromatography(hexane/acetone 95/5 to hexane/acetone 9/1) yielded the hydroxyketone J4as a clear colorless oil (481 mg, 0.75 mmol, 87%).

(3S,7S,10R,Z)-2-(3′-bromophenyl)-3-hydroxy-7-((4″-methoxybenzyl)oxy)-10-(((2′″-methoxyethoxy)methoxy)methyl)-2,8-dimethyldodec-8-en-5-one

Formula: C₃₃H₄₇BrO₇

Molar mass: 635.63

R_(f): 0.32 (hexane/acetone 8/2)

HR-MS: calculated (M+NH₄+) 652.2843. found 652.2850. calculated (M+Na⁺)657.2397. found 657.2411.

¹H NMR (500 MHz, CD₂Cl₂) 7.52 (app. t, J=1.9 Hz, 1H), 7.34 (ddd, J=7.8,2.1, 1.0 Hz, 1H), 7.31 (ddd, J=8.0, 1.9, 1.0 Hz, 1H), 7.20-7.14 (m, 3H),6.85 (app. d, 8.7 Hz, 2H), 5.18 (dd, J=10.5, 1.5 Hz, 1H), 4.72 (dd,J=10.1, 3.0 Hz, 1H), 4.64 (d, J=6.6 Hz, 1H, A part of AB-spinsystem),4.63 (d, J=6.6 Hz, 1H, B part of AB-spinsystem), 4.31 (d, J=11.1 Hz,1H), 4.124 (d, J=11.1 Hz, 1H), 4.118 (dd, J=10.5, 1.8 Hz, 1H), 3.79 (s,3H), 3.63-3.60 (m, 2H), 3.51-3.48 (m, 2H), 3.46 (dd, J=9.4, 5.8 Hz, 1H),3.37 (dd, J=9.4, 7.0 Hz, 1H), 3.31 (s, 3H), 2.85 (dd, J=15.3, 10.1 Hz,1H), 2.62-2.53 (m, 1H), 2.43 (dd, J=17.2, 1.8 Hz, 1H), 2.26 (dd, J=17.2,10.4 Hz, 1H), 2.20 (dd, J=15.3, 3.0 Hz, 1H), 1.69 (d, J=1.4 Hz, 3H),1.55-1.45 (m, 1H), 1.28 (s, 3H), 1.26 (s, 3H), 1.20-1.10 (m, 1H), 0.78(app. t, J=7.5 Hz, 3H)

¹³C NMR (125 MHz, CD₂Cl₂) δ 210.22 (Cq), 159.59 (Cq), 150.01 (Cq),135.28 (Cq), 132.56 (CH), 131.10 (Cq), 130.30 (CH), 130.06 (CH), 129.73(2×CH), 129.49 (CH), 125.90 (CH), 122.73 (Cq), 113.99 (2×CH), 95.84(CH2), 74.65 (CH), 73.51 (CH), 72.21 (CH2), 71.71 (CH2), 70.15 (CH2),67.13 (CH2), 58.98 (CH3), 55.61 (CH3), 48.03 (CH2), 46.16 (CH2), 42.16(Cq), 39.71 (CH), 25.35 (CH2), 25.12 (CH3), 23.66 (CH3), 17.97 (CH3),11.92 (CH3) 3.5 Synthesis of J7

To a solution of propionaldehyde (200 μl, 2.79 mmol, 4 eq.) in THF (5ml) was added SmI₂ (3.5 ml, 0.35 mmol, 0.5 eq., 0.1M solution in THF) at0° C. The solution colored blue, changed rapidly to green and in the endto yellow. The solution was further cooled to −20° C. and a solution ofhydroxyketone J4 (443 mg, 0.70 mmol, 1 eq.) in THF (2 ml) was addedusing a double-ended needle and the flask containing the startingmaterial was rinsed with THF (5 ml). The reaction was stirred at −20° C.for 4 hours, after which tlc-analysis (hexane/acetone 8/2) showedcomplete conversion (There was only a very subtle difference inRf-value, however, when looking at the back of the tlc-plate afterstaining with the molybdate containing reagent, the starting materialhad a red/pink shine, whereas the target material had a blue/greenshine). The reaction was quenched by adding NaHCO₃ (15 ml) and water (10ml), extracted with EtOAc (3×25 ml), dried over MgSO₄ and concentrated.

Purification using flash column chromatography (hexane/acetone87.5/12.5) delivered alcohol J7.

(3S,5R,7S,10R,Z)-2-(3′-bromophenyl)-5-hydroxy-7-((4″-methoxybenzyl)oxy)-10-(((2′″-methoxyethoxy)methoxy)methyl)-2,8-dimethyldodec-8-en-3-ylpropionate

Formula: C₃₆H₅₃BrO₈

Molar mass: 693.71

R_(f): 0.25 (hexane/acetone 8/2)

HRMS: calculated (M+NH₄+) 710.3262. found 710.3254. calculated (M+Na⁺)715.2816. found 715.2823.

¹H NMR (500 MHz, CDCl₃) δ 7.49 (app. t, J=1.9 Hz, 1H), 7.33 (ddd, J=7.8,2.0, 1.0 Hz, 1H), 7.31 (ddd, J=7.9, 2.0, 1.0 Hz, 1H), 7.20-7.14 (m, 3H),6.84 (app. d, J=8.7 Hz, 2H), 5.33 (dd, J=10.8, 1.8 Hz, 1H), 5.18 (dd,J=10.4, 1.6 Hz, 1H), 4.68 (d, J=7.0 Hz, 1H, A part of AB-spinsystem),4.67 (d, J=7.0 Hz, 1H, B part of AB-spinsystem), 4.43 (dd, J=9.1, 4.7Hz, 1H), 4.36 (d, J=11.2 Hz, 1H), 4.15 (d, J=11.2 Hz, 1H), 3.79 (s, 3H),3.67-3.64 (m, 2H), 3.55-3.52 (m, 2H), 3.52-3.46 (m, 1H), 3.47 (dd,J=9.5, 5.9 Hz, 1H), 3.39 (dd, J=9.4, 6.8 Hz, 1H), 3.36 (s, 3H),2.57-2.48 (m, 1H), 2.27 (qd, J=7.6, 3.8 Hz, 2H), 1.87 (app. dt, J=14.3,9.0 Hz, 1H), 1.67 (d, J=1.4 Hz, 3H), 1.51-1.31 (m, 4H), 1.300 (s, 3H),1.297 (s, 3H), 1.19-1.12 (m, 1H), 1.08 (app. t, J=7.6 Hz, 3H), 0.79(app. t, J=7.5 Hz, 3H)

¹³C NMR (125 MHz, CDCl₃) δ 174.87 (Cq), 159.24 (Cq), 148.91 (Cq), 135.48(Cq), 132.15 (CH), 130.79 (Cq), 129.93 (CH), 129.86 (CH), 129.47 (CH),129.37 (2×CH), 125.41 (CH), 122.55 (Cq), 113.95 (2×CH), 95.67 (CH₂),76.84 (CH), 76.01 (CH), 71.93 (CH₂), 71.41 (CH₂), 69.61 (CH₂), 66.88(CH₂), 59.17 (CH), 55.42 (CH₃), 41.90 (Cq), 40.80 (CH₂), 39.18 (CH),38.12 (CH₂), 27.91 (CH₂), 25.26 (CH₃), 25.22 (CH₂), 24.21 (CH₃), 17.87(CH₃), 11.81 (CH₃), 9.40 (CH₃)

Preparation of SmI₂ (Szostak et al., J. Org. Chem., 2012, 77 (7), pp3049-3059).

Metallic samarium (601 mg, 4 mmol, 2 eq.) was first activated bystirring dry in a flask fitted with a rubber septum for 2 days under Aratmosphere. After activation, THF (16.5 ml) was added to the metal,followed by a red solution of I₂ (508 mg, 2 mmol, 1 eq.) in THF (3.5ml). The septum was then replaced by a glass stopper and the brownreaction mixture was heated to 60° C. Stirring was continued for 24 h at60° C., after which the solution turned dark blue. The mixture wascooled down to RT and the flask was fitted with a septum again andflushed with argon. This procedure yielded a solution of SmI₂ in THFwith a concentration of 0.1 M. Upon stirring, the solution can bemaintained for a couple of days. Two hours before using it, stirring hadto be stopped.

3.6. Synthesis of J26

To a solution of alcohol J7 (811 mg, 1.17 mmol, 1 eq.) in CH₂Cl₂ (29 ml)were added molecular sieves (4 Å) (470 mg), and the suspension wasstirred at RT. After 30′ 1,8-bis(dimethylamino)naphtalene (652 mg, 3.04mmol, 2.6 eq.) and trimethyloxonium tetrafluoroborate (433 mg, 2.92mmol, 2.5 eq.) were added, respectively. The mixture was stirred for 24h, after which tlc-analysis (hexane/acetone 8/2) showed completeconversion of the starting material. The reaction mixture was filtered,rinsed with 100 ml EtOAc and washed with a solution of CuSO₄ in water(2×80 ml, 1M). The aqueous phases were combined and extracted with EtOAc(3×150 ml). All organic phases were collected, dried over MgSO₄ andconcentrated in vacuo. Flash column chromatography (hexane/EtOAc 7/3)delivered the methylether J26 (805 mg, 1.14 mmol, 97%) as a clear oil.

(3S,5R,7S,10R,Z)-2-(3′-bromophenyl)-5-methoxy-7-((4″-methoxybenzyl)oxy)-10-(((2′″-methoxyethoxy)methoxy)methyl)-2,8-dimethyldodec-8-en-3-ylpropionate

Formula: C₃₇H₅₅BrO₈

Molar mass: 707.74

R_(f): 0.31 (hexane/acetone 8/2)

¹H NMR (500 MHz, CDCl₃) δ 7.50 (app. t, J=1.9 Hz, 1H) 7.32 (app. dd,J=7.9, 1.9 Hz, 2H) 7.18-7.13 (m, 3H) 6.85 (app. d, J=8.7 Hz, 2H) 5.42(dd, J=9.8, 2.3 Hz, 1H) 5.12 (dd, J=10.5, 1.0 Hz, 1H) 4.69 (d, J=6.7 Hz,1H, A part of AB-spinsystem) 4.68 (d, J=6.7 Hz, 1H, B part ofAB-spinsystem) 4.31 (d, J=11.2 Hz, 1H) 4.19 (dd, J=10.1, 2.6 Hz, 1H)4.04 (d, J=11.3 Hz, 1H) 3.81 (s, 3H) 6.68-3.65 (m, 2H) 3.56-3.53 (m, 2H)3.47 (dd, J=9.2, 5.9 Hz, 1H) 3.41-3.36 (m, 1H) 3.38 (s, 3H) 3.23 (s, 3H)3.18-3.12 (m, 1H) 2.50-2.42 (m, 1H) 2.24 (app. q, J=7.6 Hz, 2H) 2.04(ddd, J=14.1, 10.3, 3.7 Hz, 1H) 1.67 (d, J=1.3 Hz, 3H) 1.53-1.38 (m, 3H)1.33-1.27 (m, 1H) 1.275 (s, 3H) 1.266 (s, 3H) 1.18-1.10 (m, 1H) 1.06(app. t, J=7.6 Hz, 3H) 0.77 (app. t, J=7.4 Hz, 3H)

¹³C NMR (125 MHz, CDCl₃) δ 173.88 (Cq), 159.11 (Cq), 149.03 (Cq), 136.46(Cq), 131.20 (Cq), 130.00 (CH), 129.76 (CH), 129.36 (CH), 129.22 (2×CH),125.55 (CH), 122.45 (Cq), 113.86 (2×CH), 95.66 (CH₂), 76.36 (CH),74.93(CH), 73.23 (CH), 71.93 (CH₂), 71.54 (CH₂), 69.69 (CH₂), 66.88 (CH₂),59.18 (CH₃), 56.58 (CH₃), 55.44 (CH₃), 42.04 (Cq), 39.33 (CH), 37.49(CH₂), 35.20 (CH₂), 27.90 (CH₂), 25.56 (CH₃), 25.22 (CH₂), 24.04 (CH₃),18.04 (CH₃), 11.95 (CH₃), 9.50 (CH₃),

3.7 Synthesis of S2

To a cooled (0° C.) suspension of LiAlH₄ (152 mg, 4.01 mmol, 3 eq.) inEt₂O (32 ml) was added a solution of ester J26 (946 mg, 1.34 mmol, 1eq.) in Et₂O (32 ml) using a double-ended needle. The solution wasstirred for 1 h at 0° C., upon which tlc-analysis (hexane/EtOAc 7/3)indicates full conversion of the starting material. The excess LiAlH₄was quenched by adding EtOAc (10 ml). Then, a saturated aqueous solutionof Rochelle's salt (25 ml) was added, and the resulting mixture wasstirred for 1 h at RT. The mixture was transferred to a separationfunnel containing another portion of a saturated aqueous solution ofRochelle's salt (25 ml), extracted with EtOAc (3×50 ml), dried overMgSO₄ and concentrated. Flash column chromatography (hexane/acetone 8/2)delivered alcohol S2 (871 mg, 1.34 mmol, 100%) as an oil.

(3S,5S,7S,10R,Z)-2-(3′-bromophenyl)-5-methoxy-7-((4″-methoxybenzyl)oxy)-10-(((2′″-methoxyethoxy)methoxy)methyl)-2,8-dimethyldodec-8-en-3-ol

Formula: C₃₄H₅₁BrO₇

Molar mass: 651.67

R_(f): 0.40 (hexane/acetone 7/3)

¹H NMR (500 MHz, CDCl₃) δ 7.49 (app. t, J=1.9 Hz, 1H), 7.32 (ddd, J=7.9,2.0, 1.0 Hz, 1H), 7.29 (ddd, J=7.9, 1.8, 1.0 Hz, 1H), 7.21 (app. d,J=8.8 Hz, 2H), 7.16 (app. t, J=7.9 Hz, 1H), 6.86 (app. d, J=8.7 Hz, 2H),5.17 (dd, J=10.4, 1.6 Hz, 1H), 4.70 (d, J=6.7 Hz, 1H, A part ofAB-spinsystem), 4.69 (d, J=6.7 Hz, 1H, B part of AB-spinsystem), 4.35(d, J=11.3 Hz, 1H), 4.24 (dd, J=10.0, 3.3 Hz, 1H), 4.04 (d, J=11.3 Hz,1H), 3.81 (s, 3H), 3.81-3.78 (m, 1H), 3.69-3.66 (m, 2H), 3.56-3.50 (m,3H), 3.50 (dd, J=9.4, 5.9 Hz, 1H), 3.42 (dd, J=9.5, 6.9 Hz, 1H), 3.37(s, 3H), 3.25 (s, 3H), 2.97 (bs, 1H), 2.57-2.48 (m, 1H), 2.17 (ddd,J=14.3, 10.1, 4.2 Hz, 1H), 1.71 (d, J=1.5 Hz, 3H), 1.58-1.50 (m, 1H),1.49 (ddd, J=14.3, 7.8, 3.3 Hz, 1H), 1.43 (ddd, J=14.8, 10.3, 4.6 Hz,1H), 1.34 (ddd, J=14.7, 5.9, 2.0 Hz, 1H), 1.24-1.17 (m, 1H), 1.23 (s,3H), 1.22 (s, 3H), 0.83 (app. t, J=7.5 Hz, 3H)

¹³C NMR (125 MHz, CDCl₃) δ 159.31 (Cq), 150.10 (Cq), 136.19 (Cq), 131.54(CH), 130.87 (Cq), 130.07 (CH), 129.72 (CH), 129.64 (2×CH), 129.12 (CH),125.63 (CH), 122.56 (Cq), 113.93 (2×CH), 95.69 (CH₂), 77.33 (CH), 75.88(CH), 73.08 (CH), 71.93 (CH₂), 71.47 (CH₂), 69.63 (CH₂), 66.89 (CH₂),59.18 (CH₃), 56.55 (CH₃), 55.46 (CH₃), 42.64 (Cq), 39.31 (CH), 36.26(CH₂), 33.53 (CH₂), 25.28 (CH₂), 24.53 (CH₃), 23.98 (CH₃), 18.12 (CH₃),11.87 (CH₃)

3.8 Synthesis of S3

In a pressure tube was added to a solution of alcohol S2 (873 mg, 1.34mmol, 1 eq.) in toluene (13 ml), respectively hexamethylditin (555 μl,2.68 mmol, 2 eq.), AsPh₃ (164 mg, 0.54 mmol, 0.4 eq.) andtetrakis(triphenylphosphine)palladium(0) (50 mg, 34 μmol, 3.3 mol %).The solution was heated to 70° C. and stirred for 94 h, after whichthere was clear formation of Pd-black. Although tlc-analysis(hexane/acetone 8/2) did not show complete conversion, the reaction wasquenched using a saturated aqueous solution of NaHCO₃ (15 ml) and water(15 ml). The aqueous phase was extracted with EtOAc (3×30 ml), driedover MgSO₄ and concentrated. Starting and target material were isolatedtogether using flash column chromatography (hexane/acetone 8/2). Thismixture was then put back into reaction, using the same conditions asdescribed above. Every 20 h fresh catalyst (50 mg, 34 μmol, 3.3 mol %)was added for 3 times. Quenching and workup was accomplished asdescribed. After performing flash column chromatography, the mixture ofstarting and target material was put into reaction for a 3rd time, butnow using 4 eq. of hexamethylditin, to prevent homo-coupling. Quenchingand workup proceeded exactly as described above. Flash columnchromatography (hexane/acetone 9/1) now provided the stannane S3 (804mg, 1.09 mmol, 81%) as a sticky oil.

(3S,5S,7S,10R,Z)-5-methoxy-7-((4′-methoxybenzyl)oxy)-10-(((2″-methoxyethoxy)methoxy)methyl)-2,8-dimethyl-2-(3′″-(trimethylstannyl)phenyl)dodec-8-en-3-ol

Formula: C₃₇H₆₀O₇Sn

Molar mass: 735.58

R_(f): 0.28 (hexane/acetone 8/2)

1H NMR (500 MHz, CDCl₃) δ 7.49-7.47 (m, 1H), 7.33-7.27 (m, 3H), 7.21(app. d, J=8.7 Hz, 2H) 6.85 (app. d, J=8.7 Hz, 2H), 5.17 (dd, J=10.5,1.7 Hz, 1H), 4.70 (d, J=6.7 Hz, 1H, A part of AB-spinsystem), 4.69 (d,J=6.7 Hz, 1H, B part of AB-spinsystem), 4.34 (d, J=11.3 Hz, 1H), 4.28(dd, J=9.6, 3.5 Hz, 1H), 4.06 (d, J=11.3 Hz, 1H), 3.87 (dd, J=10.1, 3.4Hz, 1H), 3.80 (s, 3H), 3.68-3.65 (m, 2H), 3.56-3.50 (m, 3H), 3.49 (dd,J=9.3, 5.8 Hz, 1H), 3.42 (dd, J=9.3, 6.8 Hz, 1H), 3.37 (s, 3H), 3.24 (s,3H), 2.66 (d, J=3.4 Hz, 1H), 2.60-2.51 (m, 1H), 2.15 (ddd, J=14.4, 9.7,4.5 Hz, 1H), 1.70 (d, J=1.4 HZ, 3H), 1.56-1.46 (m, 3H), 1.41 (ddd,J=14.4, 6.4, 2.1 Hz, 1H), 1.28 (s, 3H), 1.25 (s, 3H), 1.24-1.16 (m, 1H),0.83 (app. t, J=7.5 Hz, 3H), 0.27 (s, 9H, satellite peaks: J=27.5, 26.2Hz)

¹³C NMR (125 MHz, CDCl₃) δ 159.24 (Cq), 146.93 (Cq), 141.96 (Cq), 136.28(Cq), 133.85 (CH), 133.60 (CH), 131.44 (CH), 131.00 (Cq), 129.62 (2×CH),127.84 (CH), 126.79 (CH), 113.90 (2×CH), 95.69 (CH₂), 77.21 (CH), 76.05(CH), 73.35 (CH), 71.93 (CH₂), 71.44 (CH₂), 69.71 (CH₂), 66.88 (CH₂),59.18 (CH₃), 56.43 (CH₃), 55.43 (CH₃), 42.54 (Cq), 39.23 (CH), 36.75(CH₂), 34.04 (CH₂), 25.29 (CH₂), 24.43 (CH₃), 23.73 (CH₃), 18.33 (CH₃),11.85 (CH₃), −9.34 (3×CH₃)

3.9 Synthesis of D1

To a solution of stannane S3 (250 mg, 0.34 mmol, 1 eq.) in THF (1.7 ml)were added methyl-E-4-bromobutenoate (81 μl, 0.68 mmol, 2 eq.) andPd₂dba₃.CHCl₃ (5 mg, 5.1 μmol, 1.5 mol %) in a pressure tube and theresulting red-brown mixture was heated to 70° C., upon which it colouredyellow. Tlc-analysis (hexane/acetone 7/3) after 4 h showed completeconversion of the starting material. The reaction was then quenched witha saturated aqueous solution of NaHCO₃ (15 ml), extracted with CH₂Cl₂(4×15 ml), dried over MgSO₄ and concentrated. Flash columnchromatography (hexane/acetone 8/2) yielded the unsaturated ester D1(228 mg, 0.34 mmol, 100%) as a yellow oil.

Methyl(E)-4-(3′-((3″S,5″S,7″S,10″R,Z)-3″-hydroxy-5″-methoxy-7″-((4′″-methoxybenzyl)oxy)-10″-(((2″″-methoxyethoxy)methoxy)methyl)-2″,8″-dimethyldodec-8″-en-2″-yl)phenyl)but-2-enoate

Formula: C₃₉H₅₈O₉

Molar mass: 670.87

R_(f): 0.14 (hexane/acetone 8/2)

¹H NMR (500 MHz, CDCl3) δ 7.25-7.19 (m, 4H), 7.16-7.14 (m, 1H), 7.09(app. dt, J=15.6, 6.9 Hz, 1H), 7.00-6.97 (m, 1H), 6.85 (app. d, J=8.9Hz, 2H), 5.81 (app. dt, J=15.4, 1.7 Hz, 1H), 5.17 (app. dd, J=10.4, 1.5Hz, 1H), 4.70 (d, J=6.7 Hz, 1H, A part of AB-spinsystem), 4.69 (d, J=6.7Hz, 1H, B part of AB-spinsystem), 4.35 (d, J=11.4 Hz, 1H), 4.27 (dd,J=9.8, 3.4 Hz, 1H), 4.05 (d, J=11.3 Hz, 1H), 3.83 (ddd, J=10.3, 3.4, 2.0Hz, 1H), 3.80 (s, 3H), 3.71 (s, 3H), 3.68-3.65 (m, 2H), 3.56-3.46 (m,6H), 3.42 (dd, J=9.4, 6.8 Hz, 1H), 3.37 (s, 3H), 3.23 (s, 3H), 2.80 (d,J=3.4 Hz, 1H), 2.59-2.50 (m, 1H), 2.16 (ddd, J=14.1, 9.6, 4.6 Hz, 1H),1.70 (d, J=1.6 Hz, 3H), 1.57-1.46 (m, 2H), 1.44 (ddd, J=14.7, 10.2, 4.6Hz, 1H), 1.36 (ddd, J=14.5, 6.2, 2.0 Hz, 1H), 1.27-1.19 (m, 1H), 1.24(s, 3H), 1.23 (s, 3H), 0.83 (app. t, J=7.5 Hz, 3H)

¹³C NMR (125 MHz, CDCl₃) δ 167.06 (Cq), 159.26 (Cq), 148.08 (Cq), 148.00(CH), 137.36 (Cq), 136.25 (Cq), 131.46 (CH), 130.95 (Cq), 129.64 (2×CH),128.48 (CH), 127.29 (CH), 126.42 (CH), 125.21 (CH), 121.92 (CH), 113.90(2×CH), 95.68 (CH₂), 77.26 (CH), 76.01 (CH), 73.20 (CH), 71.92 (CH₂),71.42 (CH₂), 69.64 (CH₂), 66.88 (CH₂), 59.17 (CH₃), 56.45 (CH₃), 55.42(CH₃), 51.59 (CH₃), 42.44 (Cq), 39.24 (CH), 38.92 (CH₂), 36.53 (CH₂),33.72 (CH₂), 25.28 (CH₂), 24.45 (CH₃), 23.97 (CH₃), 18.11 (CH₃), 11.83(CH₃)

3.10 Synthesis of J70

To a solution of ester unsaturated ester D1 (111 mg, 0.18 mmol, 1 eq.)in MeOH (3.6 ml), was added NiCl₂.6H₂O (1 mg, 0.7 μmol, 4 mol %) and thered solution was cooled to 0° C. Then, NaBH₄ (14 mg, 0.36 mmol, 2 eq.)was added, gas started to evolve and the solution turned first brown,then black. After 10′, tlc-analysis (hexane/acetone 8/2) showed completeconversion of the starting material, upon which the reaction mixture wasfiltered over Celite® and concentrated. Flash column chromatography(hexane/acetone 8/2) yielded saturated ester J70 (116 mg, 0.17 mmol,96%) as a colorless oil.

Methyl4-(3′-((3″S,5″S,7″S,10″R,Z)-3″-hydroxy-5″-methoxy-7″-((4′″-methoxybenzyl)oxy)-10″-(((2″″-methoxyethoxy)methoxy)methyl)-2″,8″-dimethyldodec-8″-en-2″-yl)phenyl)-butanoate

Formula: C₃₉H₆₀O₉

Molar mass: 672.90

R_(f): 0.3 (hexane/acetone 8/2)

HRMS: calculated (M+Na⁺) 695.4130. found 695.4115.

[α]_(D): −48.2° (c=7.3 mg/ml)

¹H NMR (500 MHz, CDCl₃) δ (7.23-7.16 (m, 5H), 7.00 (app. dt, J=6.7, 1.8Hz, 1H), 6.85 (app. d, J=8.7 Hz, 2H), 5.17 (dd, J=10.5, 1.6 Hz, 1H),4.69 (d, J=6.6 Hz, 1H, A part of AB-spinsystem), 4.68 (d, J=6.6 Hz, 1H,B part of AB-spinsystem), 4.34 (d, J=11.3 Hz, 1H), 4.28 (dd, J=9.6, 3.5Hz, 1H), 4.07 (d, J=11.3 Hz, 1H), 3.85 (dd, J=9.5, 2.8 Hz, 1H), 3.80 (s,3H), 3.69-3.65 (m, 2H), 3.66 (s, 3H), 3.56-3.48 (m, 3H), 3.49 (dd,J=9.5, 5.9 Hz, 1H), 3.42 (dd, J=9.4, 6.7 Hz, 1H), 3.37 (s, 3H), 3.24 (s,3H), 2.65-2.52 (m, 3H), 2.32 (app. t, J=7.4 Hz), 2H), 2.15 (ddd, J=14.4,9.8, 4.5 Hz, 1H), 1.98-1.91 (m, 2H), 1.70 (d, J=1.4 Hz, 3H), 1.56-1.37(m, 4H), 1.27 (s, 3H), 1.25 (s, 3H), 1.24-1.18 (m, 1H), 0.84 (app. t,J=7.5 Hz, 3H)

¹³C NMR (125 MHz, CDCl₃) δ 174.05 (Cq), 159.34 (Cq), 147.70 (Cq), 141.14(Cq), 136.38 (Cq), 131.41 (CH), 131.18 (Cq), 129.56 (2×CH), 128.22 (CH),127.08 (CH), 126.11 (CH), 124.52 (CH), 113.98 (2×CH), 95.77 (CH₂), 77.26(CH), 76.10 (CH), 73.49 (CH), 72.01 (CH₂), 71.54 (CH₂), 69.76 (CH₂),66.98 (CH₂), 59.13 (CH₃), 56.41 (CH₃), 55.45 (CH₃), 51.56 (CH₃), 42.48(Cq), 39.29 (CH), 36.95 (CH₂), 35.62 (CH₂), 34.23 (CH₂), 33.67 (CH₂),26.79 (CH₂), 25.35 (CH₂), 24.54 (CH₃), 23.94 (CH₃), 18.10 (CH₃), 11.78(CH₃)

3.11 Synthesis of J71

To a solution of ester J70 (68 mg, 102 μmol, 1 eq.) in CH₂Cl₂ (5 ml) andpH 7 phosphate buffer (0.5 ml) at 0° C. was added2,3-dichloro-5,6-dicyanobenzoquinone (115 mg, 508 μmol, 5 eq.) in oneportion. The reaction mixture was stirred at 0° C. for 2 h, after whichtlc-analysis (hexane/acetone 8/2) showed complete conversion of thestarting material. The reaction was quenched using a saturated aqueoussolution of NaHCO₃ (5 ml) and water (5 ml), diluted with CH₂Cl₂ (5 ml)and the phases were separated. The aqueous phase was extracted withCH₂Cl₂ (3×10 ml), the combined organic phases were dried over MgSO₄ andconcentrated. Flash column chromatography (hexane/acetone 8/2) provideddiol J71 as a colorless oil (53 mg, 96 μmol, 96%).

Methyl-4-(3′-((3″S,5″R,7″S,10″R,Z)-3″,7″-dihydroxy-5″-methoxy-10″-(((2′″-methoxyethoxy)methoxy)methyl)-2″,8″-dimethyldodec-8″-en-2″-yl)phenyl)butanoate

Formula: C₃₁H₅₂O₈

Molar mass: 552.74

R_(f): 0.07 (hexane/acetone 8/2)

HR-MS: calculated for (M+Na+) 575.3554. found 575.3550.

[α]_(D): −15.8° (c=7.3 mg/ml)

¹H NMR (500 MHz, CDCl₃) δ 7.25-7.18 (m, 3H), 7.04-6.99 (m, 1H), 4.97(dd, J=10.1, 1.6 Hz, 1H), 4.68 (d, J=6.8 Hz, 1H, A part ofAB-spinsystem), 4.66 (d, J=6.8 Hz, 1H, B part of AB-spinsystem), 4.57(dd, J=8.7, 4.6 Hz, 1H), 3.90 (dd, J=10.2, 1.4 Hz, 1H), 3.67-3.61 (m,2H), 3.65 (s, 3H), 3.56-3.44 (m, 4H), 3.38 (s, 3H), 3.29-3.23 (m, 1H),3.28 (s, 3H), 3.00 (bs, 1H), 2.84 bs, 1H), 2.64-2.54 (m, 3H), 2.33 (app.t, J=15.2 Hz, 2H), 2.03 (ddd, J=14.4, 8.5, 6.9 Hz, 1H), 1.99-1.91 (m,2H), 1.70 (d, J=1.3 Hz, 3H), 1.62-1.38 (m, 4H), 1.33 (s, 3H), 1.32 (s,3H), 1.23-1.12 (m, 1H), 0.84 (app. t, J=7.5 Hz, 3H)

¹³C NMR (125 MHz, CDCl₃) δ 174.11 (Cq), 147.49 (Cq), 141.19 (Cq), 139.60(Cq), 130.67 (CH), 128.27 (CH), 127.02 (CH), 126.23 (CH), 124.42 (CH),95.50 (CH₂), 78.66 (CH), 76.24 (CH), 71.90 (CH₂), 71.26 (CH₂), 67.02(CH₂), 66.49 (CH), 59.15 (CH₃), 56.80 (CH₃), 51.66 (CH₃), 42.46 (Cq),39.30 (CH), 37.27 (CH₂), 35.57 (CH₂), 34.25 (CH₂), 33.61 (CH₂), 26.78(CH₂), 25.04 (CH₂), 24.51 (CH₃), 23.82 (CH₃), 18.30 (CH₃), 11.95 (CH₃)

3.12 Synthesis of J72

To a solution of diol J71 (10 mg, 18 μmol, 1 eq.) in a mixture of THF(360 μl) and water (360 μl) was added LiOH.H₂O (7 mg, 180 mmol, 10 eq.)at RT. The reaction mixture was stirred overnight. Tlc-analysis(hexane/acetone 6/4) indicated complete conversion of the startingmaterial. The reaction mixture was then poured into a saturated aqueousNH₄Cl solution (5 ml), extracted with EtOAc (5×5 ml), dried over MgSO₄and concentrated. Dry toluene (1 ml) was added to the crude product andevaporated, yielding carboxylic acid J72 (10 mg, 18 μmol, 100%) as aclear oil, which was used without further purification.

4-(3-((3S,5R,7S,10R,Z)-3,7-dihydroxy-5-methoxy-10-(((2-methoxyethoxy)methoxy)methyl)-2,8-dimethyldodec-8-en-2-yl)phenyl)butanoicacid

Formula: C₃₀H₅₀O₈

Molar mass: 538.72

R_(f): 0.55 (CH₂Cl₂/MeOH/AcOH 95/5/1)

3.13 Synthesis of J73

To a solution of the crude carboxylic acid J72 (10 mg, 18 μmol, 1 eq.)in PhMe (540 μl) was added consecutive DIPEA (24 μl, 135 μmol, 7.5 eq.)and 2,4,6-trichlorobenzoylchloride (14 μl, 90 μmol, 5 eq.) at RT. After45′ of anhydride formation, the reaction mixture was diluted with PhMe(5 ml), and sucked into a syringe. The flask was rinsed with PhMe (4.4ml in 3 times) and everything was added to the same syringe. Thesolution containing the anhydride (10 ml total volume) was then added atRT over 12 h (0.8 ml/h) to a solution of DMAP (55 mg, 450 mmol, 25 eq.)in PhMe (18 ml) using a syringe pump. 22 h after the start of theaddition, tlc-analysis (CH₂Cl₂/MeOH/AcOH 95/5/1) showed completeconversion of the starting material. The reaction was poured intodiluted HCl (20 ml, 0.1 M) and the phases were separated. The aqueousphase was extracted with EtOAc (2×30 ml), the combined organic phaseswere washed with a saturated aq. NaHCO₃-solution (25 ml), dried overMgSO₄ and concentrated. Purification using flash column chromatography(hexane/acetone 88/12) yielded the macrocyclic ester J73 as a sticky oil(8 mg, 15 μmol, 88% over 2 steps).

(3′R,7S,9R,11S,Z′)-11-hydroxy-9-methoxy-7-(3′-(2″-methoxyethoxymethoxymethyl)-1′-methylpent-1′-enyl)-12,12-dimethyl-6-oxabicyclo-[11.3.1]-heptadeca-1(17),13,15-trien-5-one

Formula: C₃₀H₄₈O₇

Molar mass: 520.71

R_(f): 0.39 (hexane/acetone 9/1)

HRMS: calculated (M+NH₄+) 538.3738. found 538.3753.

[α]_(D): −72.7° (c=7.3 mg/ml)

¹H NMR (500 MHz, CDCl₃) δ 7.28-7.23 (m, 2H), 7.06-7.01 (m, 2H), 5.52(dd, J=10.4, 1.3 Hz, 1H), 4.97 (dd, J=10.4, 1.4 Hz, 1H), 4.704 (d, J=6.9Hz, 1H, A part of AB-spinsystem), 4.695 (d, J=6.9 Hz, 1H, B part ofAB-spinsystem), 3.79-3.74 (m, 1H), 3.74-3.64 (m, 2H), 3.61-3.54 (m, 2H),3.50 (dd, J=9.4, 5.0 Hz, 1H) 3.40 (s, 3H), 3.40-3.33 (m, 2H), 3.36 (dd,J=9.4, 6.7 Hz, 1H), 3.23 (s, 3H), 2.88 (ddd, J=13.5, 8.6, 4.7 Hz, 1H),2.69-2.61 (m, 2H), 2.29-2.16 (m, 3H), 1.93 (ddd, J=17.6, 10.3, 2.9 Hz,1H), 1.89-1.79 (m, 2H), 1.66-1.50 (m, 3H), 1.57 (d, J=1.5 Hz), 1.40 (s,3H), 1.37 (s, 3H), 1.21-1.14 (m, 1H), 1.05 (ddd, J=14.8, 8.5, 2.4 Hz,1H), 0.81 (app. t, J=7.5 Hz, 3H)

¹³C NMR (125 MHz, CDCl₃) δ 171.70 (Cq), 145.84 (Cq), 139.91 (Cq), 135.04(Cq), 129.67 (CH), 128.60 (CH), 127.90 (CH), 126.57 (CH), 124.84 (CH),95.68 (CH₂), 77.03 (CH), 75.95 (CH), 72.00 (CH₂), 70.91 (CH₂), 68.37(CH), 66.75 (CH₂), 59.15 (CH₃), 56.59 (CH₃), 42.38 (Cq), 39.36 (CH),36.79 (CH₂), 35.55 (CH₂), 34.53 (CH₂), 31.49 (CH₂), 27.54 (CH₃), 25.16(CH₂), 23.47 (CH₃), 23.22 (CH₂), 18.67 (CH₃), 11.84 (CH₃)

3.14 Synthesis of J74

To a solution of the MEM-ether J73 (7 mg, 13 μmol, 1 eq.) in CH₂Cl₂ (540μl) was added n-BuSH (2 μl, 20 μmol, 1.5 eq.) and ZnBr₂ (5 mg, 20 μmol,1.5 eq.) at RT. The reaction was followed using tlc-analysis(hexane/acetone 8/2) and stirred for 25′, after which the mixture waspoured into a saturated aqueous NaHCO₃-solution (5 ml). The mixture wasextracted with CH₂Cl₂ (4×5 ml), dried over MgSO₄ and concentrated. Flashcolumn chromatography (hexane/acetone 8/2) delivered the macrocyclicdiol J74 as a white solid (5.5 mg, 12.5 μmol, 96%).

Before submitting the sample to biological testing, it was extrapurified using preparative HPLC (Luna C18 column, 50% MeCN to 100% MeCNin 30′).

(3′R,7S,9R,11S,Z′)-11-hydroxy-9-methoxy-7-(3′-hydroxymethyl-1′-methylpent-1′-enyl)-12,12-dimethyl-6-oxabicyclo-[11.3.1]-heptadeca-1(17),13,15-trien-5-one

Formula: C₂₆H₄₀O₅

Molar mass: 432.60

R_(f): 0.19 (hexane/acetone 8/2)

HRMS: calculated (M+H+) 433.2949. found 433.2958.

[α]_(D): −53.7° (c=5.6 mg/ml in CHCl₃)

¹H NMR (500 MHz, CDCl₃) δ 7.30-7.22 (m, 2H), 7.06-7.02 (m, 2H), 5.46(dd, J=10.5, 1.2 Hz, 1H), 4.91 (dd, J=10.8, 1.5 Hz, 1H), 3.76 (dd,J=8.9, 2.4 Hz, 1H), 3.63 (dd, J=10.3, 4.1 Hz, 1H), 3.46-3.40 (m, 1H),3.27 (s, 3H), 3.24 (app. t, J=10.1 Hz, 1H), 2.83 (ddd, J=13.7, 8.8, 4.6Hz, 1H), 2.67-2.58 (m, 2H), 2.27 (ddd, J=17.2, 6.8, 3.4, 1H), 2.24-2.15(m, 1H), 1.96 (ddd, J=17.2, 10.2, 3.3 Hz, 1H), 1.89 (ddd, J=15.1, 10.4,2.7 Hz, 1H), 1.88-1.80 (m, 1H), 1.65 (ddd, J=15.1, 5.8, 1.5 Hz, 1H),1.57 (d, J=1.4 Hz, 3H), 1.50 (ddd, J=14.8, 11.0, 2.4 Hz, 1H), 1.41 (s,3H), 1.41-1.34 (m, 1H), 1.38 (s, 3H), 1.11-1.04 (m, 1H), 1.01 (ddd,J=14.9, 8.9, 2.4 Hz, 1H), 0.82 (app. t, J=7.4 Hz, 3H)

¹³C NMR (125 MHz, CDCl₃) δ 173.27 (Cq), 145.74 (Cq), 139.84 (Cq), 136.10(Cq), 131.24 (CH), 128.73 (CH), 128.10 (CH), 126.56 (CH), 124.79 (CH),76.80 (CH), 75.80 (CH), 69.58 (CH), 66.85 (CH₂), 56.84 (CH₃), 42.71(CH), 42.39 (Cq), 37.04 (CH₂), 35.34 (CH₂), 34.44 (CH₂), 31.55 (CH₂),27.56 (CH₃), 24.80 (CH₂), 23.47 (CH₂), 23.07 (CH₃), 18.10 (CH₃), 12.05(CH₃)

Example 4 Synthesis of Two C₁₃-Analogues: J45 and J51 4.1 Synthesis ofTBS Ether J35

To a cooled (−78° C.) solution of alcohol J7 (1.31 g, 1.89 mmol, 1.00equivalent) in 11.3 ml CH₂Cl₂ was added 2,6-lutidine (546 μl, 4.72 mmol,2.50 equivalent), followed by TBSOTf (867 μl, 3.77 mmol, 2.00equivalent). After stirring 3 h at −78° C., 14 ml sat. aq. NaHCO₃ wasadded and stirring was continued allowing the mixture to warm to roomtemperature. The combined organic phases of the extraction with CH₂Cl₂(5×15 ml) were dried over MgSO₄ and concentrated. The orange oil (2.23g) was then purified by flash chromatography (FC) (SiO₂, CH₂Cl₂/diethylether 97/3) to provide J35 (1.41 g, 92%) as a clear yellow oil.

(3S,5R,7S,10R,Z)-2-(3′-bromophenyl)-5-(tert-butyldimethylsilyloxy)-7-(4′-methoxybenzyloxy)-10-(2′-methoxyethoxymethoxymethyl)-2,8-dimethyldodec-8-en-3-ylpropionate

Formula: C₄₂H₆₇BrO₈Si

Molar Mass: 807.97 g/mol

R_(f): 0.32 (hexane/acetone 8/2)

ESI-MS: 824.3 (M+NH₄ ⁺)

HR-MS: 824.4140 ([M+NH₄]⁺, C₄₂H₇₁NO₈Si⁺, calculated: 824.4127)

[α]_(D): −38.3 (c=0.83, CHCl₃)

[α]₃₆₅: −111.5 (c=0.83, CHCl₃)

¹H NMR (CHLOROFORM-d, 700 MHz): δ=7.50 (app t, J=1.7 Hz, 1H), 7.30-7.34(m, 2H), 7.17-7.20 (m, J=8.6 Hz, 2H), 7.16 (app t, J=7.9 Hz, 1H),6.83-6.88 (m, 2H), 5.39 (d, J=9.7 Hz, 1H), 5.11 (d, J=9.5 Hz, 1H), 4.71(d, J=6.9 Hz, 1H), 4.69 (d, J=6.7 Hz, 1H), 4.34 (d, J=11.6 Hz, 1H), 4.10(dd, J=10.9, 1.8 Hz, 1H), 4.06 (d, J=11.4 Hz, 1H), 3.78-3.82 (m(containing a singlet, 3.81), 4H), 3.67-3.70 (m, 2H), 3.55-3.58 (m, 2H),3.47 (dd, J=9.3, 5.8 Hz, 1H), 3.37-3.41 (m (containing a singlet, 3.39,4H), 2.39-2.46 (m, 1H), 2.13-2.25 (m, 2H), 2.00 (ddd, J=14.0, 11.0, 3.0Hz, 1H), 1.65 (d, J=1.1 Hz, 3H), 1.46-1.59 (m, 2H), 1.21-1.30 (m(containing two singlets, 1.28 and 1.27), 8H), 1.09-1.19 (m, 1H), 1.03(app t, J=7.6 Hz, 3H), 0.89 (s, 9H), 0.76 (app t, J=7.4 Hz, 3H), 0.09(s, 3H), 0.02 (s, 3H)

¹³C NMR (CHLOROFORM-d, 176 MHz): δ=173.8 (C), 158.8 (C), 149.0 (C),136.3 (C), 131.3 (C), 130.7 (CH), 129.8 (CH), 129.6 (CH), 129.2 (CH),128.5 (2CH), 125.4 (CH), 122.4 (C), 113.6 (2CH), 95.5 (CH₂), 76.8 (CH),73.6 (CH), 71.8 (CH₂), 71.4 (CH₂), 69.4 (CH₂), 66.9 (CH), 66.7 (CH₂),59.0 (CH₃), 55.3 (CH₃), 42.7 (C), 42.2 (CH₂), 39.2 (CH), 38.1 (CH₂),27.7 (CH₂), 26.0 (3CH₃), 25.9 (CH₃), 25.0 (CH₂), 23.4 (CH₃), 18.0 (C),17.9 (CH₃), 11.8 (CH₃), 9.3 (CH₃), −3.8 (CH₃), −4.8 (CH₃)

4.2 Synthesis of Stannane J36

To a solution of J35 (1.39 g, 1.72 mmol, 1.00 equivalent) and PPh₃ (180mg, 0.68 mmol, 0.40 equivalent) in 17.2 ml toluene was added Me₃SnSnMe₃(713 μl, 3.44 mmol, 2.00 equivalent). Pd(PPh₃)₄ (200 mg, 0.17 mmol, 0.10equivalent) was added and the yellow solution was heated at 70° C. Afterstirring for 48 h at 70° C., the reaction mixture was poured in 31 mlsat. aq. NaHCO₃ and 31 ml water, extracted with CH₂Cl₂ (3×62 ml), driedover MgSO₄ and concentrated. The residue (2.62 g, yellow oil with anorange solid) was purified by FC (SiO₂, pentane/diethyl ether 8/2) toprovide J36 (1.37 g, 89%) as a pale oil.

(3S,5R,7S,10R,Z)-5-(tert-butyldimethylsilyloxy)-7-(4′-methoxybenzyloxy)-10-(2′-methoxyethoxymethoxymethyl)-2,8-dimethyl-2-(3′-trimethylstannylphenyl)dodec-8-en-3-ylpropionate

Formula: C₄₅H₇₆O₈SiSn

Molar Mass: 891.88 g/mol

R_(f): 0.28 (pentane/diethyl ether 8/2)

ESI-MS: 910.5 [M+NH₄ ⁺]

HR-MS: 910.4696 ([M+NH₄], C₄₅H₈₀NO₈SiSn⁺, calculated: 910.4670)

[α]_(D): −32.5 (c=1.04, CHCl₃)

[α]₃₆₅: −91.4 (c=1.04, CHCl₃)

¹H NMR (CHLOROFORM-d, 500 MHz): δ=7.40-7.53 (m, 1H), 7.24-7.36 (m, 3H),7.16-7.22 (m, 2H), 6.81-6.88 (m, 2H), 5.43 (d, J=9.8 Hz, 1H), 5.11 (d,J=9.5 Hz, 1H), 4.71 (d, J=6.9 Hz, 1H), 4.69 (d, J=6.9 Hz, 1H), 4.34 (d,J=11.4 Hz, 1H), 4.12 (dd, J=10.4, 1.8 Hz, 1H), 4.07 (d, J=11.6 Hz, 1H),3.78-3.85 (m (containing a singlet, 3.81), 4H), 3.66-3.70 (m, 2H),3.52-3.58 (m, 2H), 3.47 (dd, J=9.8, 6.0 Hz, 1H), 3.35-3.42 (m(containing a singlet, 3.39), 4H), 2.40-2.49 (m, 1H), 2.11-2.18 (m, 2H),2.00 (ddd, J=14.0, 11.1, 3.2 Hz, 1H), 1.65 (d, J=1.1 Hz, 3H), 1.57-1.62(m, 1H), 1.43-1.53 (m, 1H), 1.20-1.39 (m (containing two singlets, 1.31and 1.30, 8H), 1.08-1.19 (m, 1H), 0.98 (app t, J=7.6 Hz, 3H), 0.88 (s,9H), 0.75 (app t, J=7.4 Hz, 3H), 0.28 (s, 9H), 0.08 (s, 3H), 0.01 (s,3H)

¹³C NMR (CHLOROFORM-d, 126 MHz): δ=173.7 (C), 158.8 (C), 145.8 (C),141.6 (C), 136.4 (C), 133.5 (2CH), 131.3 (C), 130.6 (CH), 128.5 (2CH),127.6 (CH), 126.7 (CH), 113.6 (2CH), 95.5 (CH2), 77.2 (CH), 73.7 (CH),71.8 (CH2), 71.4 (CH2), 69.4 (CH2), 67.1 (CH), 66.7 (CH2), 59.0 (CH3),55.2 (CH3), 42.7 (CH2), 42.1 (C), 39.2 (CH), 38.1 (CH2), 27.7 (CH2),26.1 (3CH3), 25.6 (CH3), 25.0 (CH2), 23.7 (CH3), 18.0 (C), 17.9 (CH3),11.8 (CH3), 9.3 (CH3), −3.8 (CH3), −4.8 (CH3), −9.6 (3CH3)

4.3 Synthesis of Ester J37

To a solution of J36 (1.34 g, 1.50 mmol, 1.00 equivalent) and(E)-methyl-4-bromo-but-2-enoate (360 μl, 3.01 mmol, 2.00 equivalent) in7.52 ml THF was added Pd(dba)₃.CHCl₃ (23.3 mg, 0.02 mmol, 0.015equivalent). The red solution was heated at 70° C. to become a clearyellow solution. After stirring for 5 h at 70° C., the reaction mixturewas poured in 36 ml sat. aq. NaHCO₃, extracted with CH₂Cl₂ (3×36 ml),dried over MgSO₄ and concentrated. The yellow oil (2.62 g) was purifiedby FC (SiO₂, pentane/diethyl ether 6/4) to afford J37 (1.06 g, 89%) as apale yellow oil.

(3″S,5″R,7″S,10″R,Z)-methyl4-(3′-(5″-(tert-butyldimethylsilyloxy)-7″-(4′″-methoxybenzyloxy)-10″-(2′″-methoxyethoxymethoxymethyl)-2″,8″-dimethyl-3″-(propionyloxy)dodec-8″-en-2″-yl)phenyl)but-2-enoate

Formula: C₄₇H₇₄O₁₀Si

Molar Mass: 827.17 g/mol

R_(f): 0.25 (pentane/diethyl ether 6/4)

ESI-MS: 844.4 [M+NH₄ ⁺]

HR-MS: 844.5398 ([M+NH₄], C₄₇H₇₈NO₁₀Si⁺, calculated: 844.5390), 849.4953([M+Na]⁺, C₄₇H₇₄NaO₁₀Si⁺, calculated: 849.4944)

[α]_(D): −35.8 (c=0.70, CHCl₃)

[α]₃₆₅: −101.2 (c=0.70, CHCl₃)

¹H NMR (CHLOROFORM-d, 300 MHz): δ=7.15-7.26 (m, 5H), 7.09 (dt, J=15.4,7.0 Hz, 1H), 6.96-7.02 (m, 1H), 6.81-6.91 (m, 2H), 5.82 (dt, J=15.5, 1.6Hz, 1H), 5.44 (d, J=9.6 Hz, 1H), 5.10 (dd, J=10.2, 0.9 Hz, 1H),4.67-4.73 (m, 2H), 4.34 (d, J=11.5 Hz, 1H), 4.11 (dd, J=10.5, 1.5 Hz,1H), 4.07 (d, J=11.5 Hz, 1H), 3.79-3.83 (m (containing a singlet), 3.81,4H), 3.71 (s, 3H), 3.65-3.70 (m, 2H), 3.53-3.59 (m, 2H), 3.35-3.52 (m(containing a singlet, 3.39), 7H), 2.38-2.50 (m, 1H), 2.12-2.22 (m, 2H),1.99 (ddd, J=13.9, 11.1, 3.0 Hz, 1H), 1.65 (d, J=1.1 Hz, 3H), 1.42-1.59(m, 2H), 1.17-1.34 (m (containing two singlets, 1.29 and 1.27), 8H),1.08-1.17 (m, 1H), 1.01 (app t, J=7.6 Hz, 3H), 0.88 (s, 9H), 0.76 (appt, J=7.4 Hz, 3H), 0.08 (s, 3H), 0.01 (s, 3H)

¹³C NMR (CHLOROFORM-d, 75 MHz): δ=173.8 (C), 167.6 (C), 147.7 (CH),147.1 (C), 137.2 (C), 136.4 (C), 131.3 (C), 130.6 (CH), 128.5 (2CH),128.3 (CH), 127.0 (CH), 126.4 (CH), 125.0 (CH), 121.8 (CH), 121.7 (C),113.6 (2CH), 95.5 (CH₂), 77.1 (CH), 73.6 (CH), 71.8 (CH₂), 71.4 (CH₂),69.4 (CH₂), 66.9 (CH), 66.7 (CH₂), 59.0 (CH₃), 55.2 (CH₃), 51.4 (CH₃),42.7 (C), 42.0 (CH₂), 39.2 (CH), 38.7 (CH₂), 38.0 (CH₂), 27.7 (CH₂),26.0 (4CH₃), 25.0 (CH₂), 23.2 (CH₃), 18.0 (C), 17.9 (CH₃), 11.8 (CH₃),9.3 (CH₃)-3.8 (CH₃), −4.9 (CH₃)

4.4 Synthesis of Diol J38

A solution of AD-mix-β 4.44 g), methanesulfonamide (235 mg, 2.47 mmol,2.00 equivalent) and NaHCO₃ (995 mg, 11.8 mmol, 9.60 equivalent) in 1.33ml t-BuOH and 12.3 ml H₂O was stirred for 10 min. A solution of J37 in 5ml t-BuOH was added dropwise and the flask was rinsed with t-BuOH (3×2ml). The greenish reaction mixture was vigorously stirred at roomtemperature for 41 h. 43 ml sat. aq. NaS₂O₃ and 20 ml water was addedand stirred for 1 h to become a clear solution followed by extractingwith CH₂Cl₂ (5×250 ml), drying over MgSO₄ and concentrating. The residue(pale oil with white solid) was purified by FC (SiO₂, pentane/acetone75/25) to afford J38 (1.02 g, 96%) as a pale oil.

(2S,3R,3″S,5″R,7″S,10″R,Z)-methyl-4-(3′-(5″-(tert-butyldimethylsilyloxy)-7″-(4′″-methoxybenzyloxy)-10″-(2′″-methoxyethoxymethoxymethyl)-2″,8″-dimethyl-3″-(propionyloxy)dodec-8″-en-2″-yl)phenyl)-2,3-dihydroxybutanoate

Formula: C₄₇H₇₆O₁₂Si

Molar Mass: 861.18 g/mol

R_(f): 0.28 (pentane/acetone 75/25)

ESI-MS: 878.4 [M+NH₄ ⁺]

HR-MS: 878.5453 ([M+NH₄], C₄₇H₈₀NO₁₂Si⁺, calculated: 878.5444), 883.5004([M+Na]⁺, C₄₇H₇₆NaO₁₂Si⁺, calculated: 884.4998)

[α]_(D): −17.7 (c=0.90, CHCl₃)

[α]₃₆₅: −36.5 (c=0.90, CHCl₃)

¹H NMR (CHLOROFORM-d, 500 MHz): δ=7.26-7.28 (m, 1H), 7.21-7.24 (m, 2H),7.16-7.20 (m, 2H), 7.04-7.12 (m, 1H), 6.82-6.87 (m, 2H), 5.41 (d, J=9.8Hz, 1H), 5.11 (d, J=10.1 Hz, 1H), 4.71 (d, J=6.7 Hz, 1H), 4.69 (d, J=6.7Hz, 1H), 4.34 (d, J=11.4 Hz, 1H), 4.09-4.17 (m, 3H), 4.07 (d, J=11.6 Hz,1H), 3.77-3.83 (m (containing two singlets, 3.81 and 3.80), 7H),3.67-3.70 (m, 2H), 3.54-3.58 (m, 2H), 3.45-3.50 (m, 1H), 3.34-3.44 (m(containing a singlet, 3.39), 4H), 2.86-2.98 (m, 2H), 2.41-2.51 (m, 1H),1.95-2.10 (m, 3H), 1.66 (d, J=0.9 Hz, 3H), 1.56-1.64 (m, 1H), 1.46-1.55(s, 1H), 1.38-1.45 (m, 1H), 1.20-1.37 (m (containing two singlets, 1.30and 1.29), 7H), 1.10-1.19 (m, 1H), 0.83-0.96 (m, 12H), 0.79 (app. t,J=7.3 Hz, 3H), 0.08 (s, 3H), 0.03 (s, 3H)

¹³C NMR (CHLOROFORM-d, 126 MHz): δ=173.8 (2C), 158.8 (C), 146.9 (C),137.1 (C), 136.4 (C), 131.2 (C), 130.6 (CH), 128.5 (2CH), 128.3 (CH),128.2 (CH), 126.9 (CH), 124.9 (CH), 113.6 (2CH), 95.5 (CH₂), 77.4 (CH),73.6 (CH), 73.4 (CH), 72.6 (CH), 71.8 (CH₂), 71.4 (CH₂), 69.4 (CH₂),67.0 (CH), 66.7 (CH₂), 59.0 (CH₃), 55.3 (CH₃), 52.6 (CH₃), 42.7 (CH₂),41.9 (C), 40.5 (CH₂), 39.2 (CH), 37.9 (CH₂), 27.6 (CH₂), 26.0 (3CH₃),25.0 (CH₂), 24.4 (CH₃), 24.3 (CH₃), 18.0 (C), 17.9 (CH₃), 11.8 (CH₃),9.2 (CH₃), −3.8 (CH₃), −4.9 (CH₃)

4.5 Synthesis of MOM Ether J39

To a solution of diol J38 (955 mg, 1.11 mmol, 1.00 equivalent) in 5.5 mlCH₂Cl₂ was added DIPEA (1.06 ml, 6.10 mmol, 5.50 equivalent), followedby adding freshly prepared MOM-Cl (2.252M CH₂Cl₂ and methyl acetate,2.46 ml, 5.54 mmol, 5.00 equivalent) at 0° C. After stirring 17 h at 40°C., 14 ml water was added and the mixture was extracted with diethylether (3×10 ml). The combined organic phases were dried over MgSO₄ andconcentrated. The orange oil (1.25 g) was then purified by FC (SiO₂,hexane/ethyl acetate 55/45) to provide J39 (1.03 g, 98%) as a clear oil.

(2S,3R,3″S,5″R,7″SI,10″R,Z)-)-methyl-4-(3′-(5″-tert-butyldimethylsilyloxy)-7″-(4′″-methoxybenzyloxy)-10″-(2′″-methoxyethoxymethoxy)ethyl)-2″,8″-dimethyl-3″-(propionyloxydodec-8″-en-2″-yl)phenyl)-2,3-bis(methoxymethoxy)butanoate

Formula: C₅₁H₈₄O₁₄Si

Molar Mass: 949.29 g/mol

R_(f): 0.19 (hexane/acetone 8/2)

ESI-MS: 966.5 [M+NH₄ ⁺]

HR-MS: 966.5999 ([M+NH₄], C₅₁H₈₈NO₁₄Si⁺, calculated: 966.5981), 971.5533([M+Na]⁺, C₅₁H₈₄NaO₁₄Si⁺, calculated: 971.5523)

[α]_(D): −36.7 (c=0.90, CHCl₃)

[α]₃₆₅: −91.0 (c=0.90, CHCl₃)

1H NMR (CHLOROFORM-d, 500 MHz): δ=7.21-7.26 (m, 3H), 7.17-7.21 (m, 2H),7.09 (app. dt, J=7.2, 1.4 Hz, 1H), 6.83-6.87 (m, 2H), 5.45 (d, J=9.9 Hz,1H), 5.11 (d, J=9.6 Hz, 1H), 4.77 (d, J=7.0 Hz, 1H), 4.65-4.74 (m, 3H),4.51 (d, J=7.2 Hz, 1H), 4.39 (d, J=7.2 Hz, 1H), 4.34 (d, J=11.4 Hz, 1H),4.21 (ddd, J=7.8, 6.4, 2.9 Hz, 1H), 4.16 (d, J=3.1 Hz, 1H), 4.11 (dd,J=10.5, 1.8 Hz, 1H), 4.07 (d, J=11.4 Hz, 1H), 3.77-3.84 (m (containing asinglet, 3.81), 4H), 3.75 (s, 3H), 3.67-3.70 (m, 2H), 3.54-3.57 (m, 2H),3.44-3.49 (m (containing a singlet, 3.46), 4H), 3.36-3.42 (m (containinga singlet, 3.39), 4H), 3.14 (s, 3H), 2.98 (dd, J=13.7, 7.8 Hz, 1H), 2.93(dd, J=13.7, 6.3 Hz, 1H), 2.41-2.50 (m, 1H), 2.11-2.22 (m, 2H), 1.99(ddd, J=13.9, 10.8, 2.9 Hz, 1H), 1.65 (d, J=1.2 Hz, 3H), 1.54-1.61 (m,1H), 1.46-1.52 (m, 1H), 1.18-1.32 (m (containing two singlets, 1.29 and1.26), 8H), 1.09-1.18 (m, 1H), 1.00 (app. t, J=7.6 Hz, 3H), 0.88 (s,9H), 0.77 (app. t, J=7.4 Hz, 3H), 0.09 (s, 3H), 0.01 ppm (s, 3H)

¹³C NMR (CHLOROFORM-d, 126 MHz): δ=173.8 (C), 171.1 (C), 158.8 (C),146.8 (C), 137.3 (C), 136.3 (C), 131.3 (C), 130.6 (CH), 128.5 (2CH),128.2 (CH), 127.7 (CH), 127.0 (CH), 125.0 (CH), 113.6 (2CH), 97.0 (CH₂),96.6 (CH₂), 95.5 (CH₂), 79.3 (CH), 77.1 (CH), 77.1 (CH), 73.7 (CH), 71.8(CH₂), 71.3 (CH₂), 69.4 (CH₂), 66.9 (CH), 66.7 (CH₂), 59.0 (CH₃), 56.5(CH₃), 55.7 (CH₃), 55.2 (CH₃), 51.9 (CH₃), 42.8 (CH₂), 42.0 (C), 39.2(CH), 38.0 (CH₂), 37.9 (CH₂), 27.7 (CH₂), 26.2 (CH₃), 26.0 (3CH₃), 25.0(CH₂), 23.1 (CH₃), 18.0 (C), 17.9 (CH₃), 11.8 (CH₃), 9.3 (CH₃), −3.8(CH₃), −4.8 (CH₃)

4.6 Synthesis of Alcohol J40

To a stirring solution of J39 (522 mg, 0.55 mmol, 1.00 equivalent) in 22ml CH₂Cl₂ and 2.2 ml pH 7 buffer was added2,3-dichloro-5,6-dicyano-1,4-benzoquinone (500 mg, 2.20 mmol, 4.00equivalent) in four portions over 90 min. After stirring theblack/brownish/greenish mixture for an extra 30 min, 20 ml sat. aq.NaHCO₃ was added. The mixture was extracted with CH₂Cl₂ (5×50 ml), driedover MgSO₄ and concentrated. The residue was then purified by FC (SiO₂,CH₂Cl₂/diethyl ether 97/3) to provide J40 (1.41 g, 97%) as a pale oil.

(2S,3R,3″S,5″R,7″S,10″R,Z)-methyl4-(3″-(5-tert-butyldimethylsilyloxy)-7″-hydroxy-10″-(2′″-methoxyethoxymethoxymethyl)-2″,8″-dimethyl-3″-(propionyloxy)dodec-8″-en-2″-yl)phenyl)-2,3-bis(methoxymethoxy)butanoate

Formula: C₄₃H₇₆O₁₃Si

Molar Mass: 829.14 g/mol

R_(f): 0.18 (hexane/ethyl acetate 1/1)

IR: 2955, 2885, 1739, 1595, 1466, 1363, 1253, 1201, 1181, 1153, 1100,1049, 913, 840, 776, 734

ESI-MS: 846.4 [M+NH₄ ⁺]

HR-MS: 846.5379 ([M+NH₄]⁺, C₄₃H₈₀NO₁₃Si⁺, calculated: 966.5393),851.4960 ([M+Na]⁺, C₄₃H₇₆NaO₁₃Si⁺, calculated: 851.4947)

[α]_(D): −21.2 (c=0.68, CHCl₃)

[α]₃₆₅: −40.8 (c=0.68, CHCl₃)

¹H NMR (CHLOROFORM-d, 500 MHz): δ=7.20-7.26 (m, 3H), 7.10 (app. dt,J=7.2, 1.7 Hz, 1H), 5.38 (d, J=8.9 Hz, 1H), 4.93 (d, J=9.2 Hz, 1H), 4.77(d, J=7.0 Hz, 1H), 4.72 (d, J=7.0 Hz, 1H), 4.69 (d, J=6.9 Hz, 1H), 4.67(d, J=6.7 Hz, 1H), 4.51 (d, J=7.2 Hz, 1H), 4.44 (dd, J=9.6, 2.6 Hz, 1H),4.40 (d, J=7.0 Hz, 1H), 4.22 (ddd, J=7.8, 6.4, 2.9 Hz, 1H), 4.16 (d,J=2.9 Hz, 1H), 3.76 (s, 3H), 3.68-3.74 (m, 1H), 3.62-3.66 (m, 2H),3.50-3.57 (m, 3H), 3.47 (s, 3H), 3.39 (s, 3H), 3.27 (app. t, J=8.9 Hz,1H), 3.16 (s, 3H), 2.99 (dd, J=13.7, 7.8 Hz, 1H), 2.94 (dd, J=13.6, 6.3Hz, 1H), 2.70 (br. s, 1H), 2.50-2.61 (m, 1H), 2.25-2.31 (m, 2H), 1.76(ddd, J=14.2, 9.3, 4.9 Hz, 1H), 1.67 (d, J=1.2 Hz, 3H), 1.56 (ddd,J=13.9, 9.6, 3.8 Hz, 1H), 1.36-1.51 (m, 3H), 1.31 (s, 3H), 1.27 (s, 3H),1.16-1.24 (m, 1H), 1.10 (app. t, J=7.6 Hz, 3H), 0.82-0.88 (m (containinga singlet, 0.86), 12H), 0.02 (s, 3H), −0.01 (s, 3H)

¹³C NMR (CHLOROFORM-d, 126 MHz): δ=173.9 (C), 171.1 (C), 146.7 (C),139.6 (C), 137.4 (C), 129.9 (CH), 128.2 (CH), 127.7 (CH), 127.1 (CH),125.0 (CH), 97.0 (CH₂), 96.6 (CH₂), 95.3 (CH₂), 79.4 (CH), 77.2 (CH),77.1 (CH), 71.7 (CH₂), 71.0 (CH₂), 68.7 (CH), 66.8 (CH₂), 66.0 (CH),59.0 (CH₃), 56.5 (CH₃), 55.8 (CH₃), 51.9 (CH₃), 42.2 (CH₂), 42.0 (C),39.1 (CH), 38.6 (CH₂), 37.9 (CH₂), 27.9 (CH₂), 26.2 (CH₃), 25.9 (3CH₃),24.9 (CH₂), 22.9 (CH₃), 18.3 (CH₃), 17.9 (C), 11.7 (CH₃), 9.3 (CH₃),−4.3 (CH₃), −4.7 (CH₃)

4.7 Synthesis of Carboxylic Acid J41

To a cooled (0° C.) solution of J40 (208 mg, 0.25 mmol, 1.00 equivalent)in 5.6 ml THF and 1.9 ml water was added LiOH.H₂O (105 mg, 2.51 mmol,10.0 equivalent) in one portion. After stirring 23 h at roomtemperature, the mixture was poured in 15 ml sat. aq. NH₄Cl, extractedwith ethyl acetate (5×15 ml), dried over MgSO₄ and concentrated. Theresidue was dissolved in 10 ml toluene and concentrated to afford crudeJ41 (217 mg) as a yellow, viscous oil. The carboxylic acid was usedwithout further purification.

(2S,3R,3′S,5″R,7″S,10″R,Z)-4-(3′-(5″-(tert-butyldimethylsilyloxy)-7″-hydroxy-10″-(2-methoxyethoxymethoxymethyl)-2″,8″-dimethyl-3″-(propionyloxy)dodec-8″-en-2″-yl)phenyl)-2,3-bis(methoxymethoxy)butanoicacid

Formula: C₄₂H₇₄O₁₃Si

Molar Mass: 815.11 g/mol

R_(f): 0.27 (pentane/acetone/acetic acid 69.5/30/0.5)

ESI-MS: 832.4 [M+NH₄ ⁺]

HR-MS: 832.5239 ([M+NH₄], C₄₂H₇₈NO₁₃Si⁺, calculated: 832.5237), 837.4793([M+Na]⁺, C₄₂H₇₄NaO₁₃Si⁺, calculated: 837.4791)

4.8 Synthesis of Alcohol J42

To a cooled (0° C.) solution of crude J41 (217 mg, 0.25 mmol, 1.00equivalent) in 5 ml diethyl ether and methanol (40.6 μl, 1.00 mmol, 4equivalent) was added LiBH₄ (2M in THF, 502 μl, 1.00 mmol, 4.00equivalent). After stirring for 15 min, the mixture was allowed to warmto room temperature and stirring was continued for 26 h (LC-MS analysisshowed a 70% conversion) before an extra portion of LiBH₄ (2M in THF,251 μl, 0.50 mmol, 2.00 equivalent) was added. After stirring an extra19 h, the mixture was cooled (0° C.), diluted with 10 ml ethyl acetateand quenched with 125 ml sat. aq. NH₄Cl. The mixture was extracted withethyl acetate (5×125 ml), dried over MgSO₄ and concentrated. The residuewas dissolved in 1.5 ml toluene and concentrated to afford crude J42(255 mg) as a yellow, viscous oil. The seco-acid was used withoutfurther purification (LC-MS analysis showed a 96.5% conversion of thestarting material).

(2S,3R,3″S,5″R,7″S,10″R,Z)-4-(3′-(5″-(tert-butyldimethylsilyloxy)-3″,7″-dihydroxy-10″-(2′″-methoxyethoxymethoxymethyl)-2″,8″-dimethyldodec-8″-en-2″-yl)phenyl)-2,3-bis(methoxymethoxy)butanoicacid

Formula: C₃₉H₇₀O₁₂Si

Molar Mass: 759.05 g/mol

R_(f): 0.21 (pentane/acetone/acetic acid 69.5/30/0.5)

ESI-MS: 757.3 [M−H⁺]

HR-MS: 757.4557 ([M−H⁺], C₃₉H₆₉O₁₃Si⁺, calculated: 757.4564)

4.9 Synthesis of Lactone J43

To a cooled (0° C.) solution of crude J42 (255 mg, 0.25 mmol, 1.00equivalent) in 9.5 ml THF was dropwise added DIPEA (328 μl, 1.88 mmol,7.50 equivalent) and 2,4,6-trichlorobenzoyl chloride (198 μl, 1.26 mmol,5.00 equivalent). The yellow mixture was stirred for 4 h at roomtemperature and THF was removed in vacuo. The residue was dissolved in50 ml toluene and added to a solution of DMAP (766 mg, 6.27 mmol, 25equivalent) in 326 ml toluene at room temperature over a period of 14 hvia syringe pump. After further stirring for 31 h, the mixture waspoured in 300 ml sat. aq. NaHCO₃, separated and extracted with ethylacetate (4×300 ml). The combined organic phases were dried over MgSO₄and concentrated in vacuo. The residue (yellow oil with white crystals)was then purified by FC (SiO₂, hexane/ethyl acetate 6/4) to provide J43(83.5 mg, 45% over three steps) as a pale yellow oil.

(3R,3′R,4S,7S,9R,11S,Z)-9-(tert-butyldimethylsilyloxy)-11-hydroxy-7-(3′-(2″-methoxyethoxymethoxymethyl)-1′-methylpent-1′-enyl)-2,3-bis(methoxymethoxy)-12,12-dimethyl-6-oxabicyclo[11.3.1]heptadeca-1(17),13,15-trien-5-one

Formula: C₃₉H₆₈O₁₁Si

Molar Mass: 741.04 g/mol

R_(f): 0.20 (hexane/ethyl acetate 6/4)

IR: 3008, 2951, 2930, 2894, 1749, 1468, 1442, 1364, 1254, 1217, 1150,1124, 1103, 1049, 1031, 944, 838, 758, 708, 667

ESI-MS: 758.4 [M+NH₄ ⁺]

HR-MS: 758.4892 ([M+NH₄]⁺, C₃₉H₇₂NO₁₁Si⁺, calculated: 758.4869),799.4645 ([M+CH₃COO⁺], C₄₁H₇₁O₁₃Si⁺, calculated: 799.4669)

[α]_(D): −32.6 (c=1.20, CHCl₃)

1H NMR (CHLOROFORM-d, 500 MHz): δ=7.34 (br. s, 1H), 7.29-7.24 (m, 2H),7.10 (app. dt, J=6.7, 1.5 Hz, 1H), 5.51 (dd, J=10.8, 3.1 Hz, 1H), 5.02(d, J=9.9 Hz, 1H), 4.82 (d, J=7.3 Hz, 2H), 4.81 (d, J=7.3 Hz, 1H),4.64-4.70 (m, 3H), 4.59 (d, J=6.7 Hz, 1H), 4.28 (ddd, J=10.8, 3.8, 1.5Hz, 1H), 3.94 (app. t, J=8.9 Hz, 1H), 3.64-3.70 (m, 2H), 3.62 (d, J=1.8Hz, 1H), 3.58 (dd, J=9.4, 4.5 Hz, 1H), 3.52-3.56 (m, 2H), 3.49 (s, 3H),3.40-3.46 (m (containing a singlet, 3.42), 5H), 3.39 (s, 3H), 3.25 (dd,J=13.1, 4.1 Hz, 1H), 2.95 (dd, J=13.2, 11.2 Hz, 1H), 2.47-2.59 (m, 1H),1.93-2.13 (m, 1H), 1.89 (d, J=11.0 Hz, 1H), 1.86 (d, J=11.3 Hz, 1H),1.53-1.72 (m (containing a doublet, 1.63), 6H), 1.45 (s, 3H), 1.40 (s,3H), 1.31 (ddd, J=14.3, 9.5, 1.8 Hz, 2H), 1.15-1.24 (m, 1H), 0.82 (app.t, J=7.5 Hz, 3H), 0.76 (s, 9H), 0.00 (s, 3H), −0.01 (s, 3H)

¹³C NMR (CHLOROFORM-d, 126 MHz): δ=169.5 (C), 146.4 (C), 136.9 (C),133.8 (C), 129.6 (CH), 128.7 (CH), 128.0 (CH), 128.0 (CH), 125.0 (CH),96.8 (CH₂), 96.8 (CH₂), 95.6 (CH₂), 80.3 (CH), 77.3 (CH), 75.1 (CH),71.8 (CH₂), 71.7 (CH), 70.2 (CH₂), 66.6 (CH₂), 66.5 (CH), 59.0 (CH₃),56.5 (CH₃), 55.8 (CH₃), 42.4 (C), 40.6 (CH₂), 40.0 (CH₂), 39.4 (CH),38.3 (CH₂), 26.1 (CH₃), 25.7 (3CH₃), 24.9 (CH₂), 22.2 (CH₃), 18.2 (C),17.7 (CH₃), 11.7 (CH₃), −4.1 (CH₃), −5.0 (CH₃)

4.10 Synthesis of Analogue J45

To a cooled (0° C.) solution of J43 (9 mg, 12.1 μmol, 1.00 equivalent)in 3.6 ml THF was added aqueous HCl (4M, 3.6 ml). After stirring 15 h atroom temperature, the mixture was diluted with 2 ml ethyl acetate,neutralized to pH 8 with sat. aq. NaHCO₃ and extracted with ethylacetate (5×25 ml). The combined organic extracts were dried over MgSO₄and concentrated in vacuo. The clear oil (16 mg) was purified by FC(SiO₂, hexane/acetone 6/4) to provide J45 (6 mg) as a white solid. HP-LCpurification (LUNA C18, 0% to 100% acetonitrile in aqueous 5 nM ammoniumacetate in 30 min) provided J45 (2.8 mg, 51%) as a white solid.

(3R,3′R,4S,7S,9R,11S,Z)-3,4,9,11-tetrahydroxy-7-(3′-hydroxymethyl-1′-methylpent-1′-enyl)-12,12-dimethyl-6-oxabicyclo[11.3.1]heptadeca-1(17),13,15-trien-5-one

Formula: C₂₅H₃₈O₇

Molar Mass: 450.57 g/mol

R_(f): 0.08 (hexane/acetone 6/4)

ESI-MS: 451.2 [M+H⁺], 433.2 [M−H₂O+H⁺]

1H NMR (CHLOROFORM-d, 500 MHz): δ=7.35 (br. s, 1H), 7.16-7.26 (m, 2H),7.05 (br. d, J=7.0 Hz, 1H), 5.48 (dd, J=10.4, 2.7 Hz, 1H), 4.93 (d,J=10.4 Hz, 1H), 4.34 (dd, J=11.7, 4.5 Hz, 1H), 4.14 (app. t, J=8.4 Hz,1H), 3.43-3.52 (m, 3H), 3.14-3.30 (m, 2H), 2.74 (app. t, J=12.3 Hz, 1H),2.26-2.67 (m, 6H), 1.76-1.91 (m, 2H), 1.57-1.71 (m (containing asinglet, 1.67), 4H), 1.50 (s, 3H), 1.24-1.44 (m (containing a singlet,1.36), 5H), 1.04-1.17 (m, 1H), 0.83 ppm (app. t, J=7.4 Hz, 3H)

¹³C NMR (CHLOROFORM-d, 126 MHz): δ=174.3 (C), 146.8 (C), 137.0 (C),136.7 (C), 129.9 (CH), 129.0 (CH), 128.5 (CH), 127.8 (CH), 124.6 (CH),76.1 (CH), 74.3 (CH), 73.9 (CH), 69.0 (CH), 67.0 (CH₂), 65.5 (CH), 42.7(CH), 42.5 (C), 40.7 (CH₂), 39.6 (CH₂), 38.9 (CH₂), 26.4 (CH₃), 24.5(CH₂), 20.3 (CH₃), 17.7 (CH₃), 11.9 (CH₃)

4.11 Synthesis of Alcohol J49

To a cooled (0° C.) solution of J43 (15 mg, 20.2 μmol, 1.00 equivalent)in 405 μl THF was added TBAF (1M in THF, 41 μl, 40.5 μmol, 2equivalent). After stirring 2 h at room temperature, the mixture wasconcentrated and purified by FC (SiO₂, pentane/acetone 8/2) to provideJ49 (7.7 mg, 61%) as a yellow oil.

(3R,3′R,4S,7S,9R,11S,Z)-9,11-dihydroxy-7-(3′-(2″-methoxyethoxymethoxymethyl)-1′-methylpent-1′-enyl)-2,3-bis(methoxymethoxy)-12,12-dimethyl-6-oxabicyclo[11.3.1]heptadeca-1(17),13,15-trien-5-one

Formula: C₃₃H₅₄O₁₁

Molar Mass: 626.78 g/mol

R_(f): 0.38 (pentane/acetone 7/3)

ESI-MS: 644.3 [M+NH₄ ⁺]

¹H NMR (CHLOROFORM-d, 300 MHz): δ=7.38 (app. dt, J=7.9, 1.4 Hz, 1H),7.21-7.32 (m, 2H), 7.16 (app. dt, J=7.5, 1.4 Hz, 1H), 5.46 (dd, J=10.2,2.3 Hz, 1H), 4.99 (d, J=9.4 Hz, 1H), 4.89 (d, J=7.2 Hz, 1H), 4.84 (d,J=7.0 Hz, 1H), 4.67-4.73 (m, 2H), 4.64 (d, J=6.8 Hz, 1H), 4.61 (d, J=6.6Hz, 1H), 4.17 (ddd, J=10.0, 5.3, 2.6 Hz, 1H), 4.01 (d, J=5.3 Hz, 1H),3.80 (dd, J=5.9, 4.2 Hz, 1H), 3.61-3.75 (m, 2H), 3.35-3.59 (m(containing three singlets, 3.47, 3.42 and 3.39), 14H), 3.18 (dd,J=14.1, 2.4 Hz, 1H), 2.93 (dd, J=14.1, 10.2 Hz, 1H), 2.48-2.65 (m, 1H),1.89 (ddd, J=14.5, 10.4, 2.4 Hz, 1H), 1.53-1.69 (m (containing adoublet, 1.62), 5H), 1.32-1.53 (m (containing two singlets, 1.44 and1.43), 8H), 1.06-1.23 (m, 1H), 0.81 (app. t, J=7.5 Hz, 3H)

¹³C NMR (CHLOROFORM-d, 75 MHz): δ=169.2 (C), 146.1 (C), 137.1 (C), 134.1(C), 130.2 (CH), 128.5 (CH), 127.9 (CH), 127.6 (CH), 125.5 (CH), 96.6(CH₂), 96.0 (CH₂), 95.6 (CH₂), 79.2 (CH), 78.0 (CH), 77.5 (CH), 71.8(CH₂), 70.9 (CH₂), 70.8 (CH), 66.6 (CH₂), 65.9 (CH), 59.0 (CH₃), 56.1(CH₃), 55.8 (CH₃), 42.3 (C), 41.3 (CH₂), 39.7 (CH), 38.7 (CH₂), 37.8(CH₂), 26.6 (CH₃), 24.9 (CH₂), 24.2 (CH₃), 18.1 (CH₃), 11.9 (CH₃) ppm

4.12 Synthesis of Ketone J50

To a cooled (0° C.) solution of J49 (5.3 mg, 8.46 μmol, 1.00 equivalent)in 250 μl CH₂Cl₂ was added pyridine (3.4 μl, 42.3 μmom, 5.00 equivalent)and Dess-Martin periodinane (4 mg, 9.31 μmol, 1.10 equivalent). Afterstirring 40 min at room temperature, 1 ml sat. aq. Na₂S₂O₃ was added andstirred further for 90 min. The two-phase mixture was separated,extracted with ethyl acetate (3×2 ml), dried over MgSO₄ andconcentrated. The ketone was used without further purification.

(3R,3′R,4S,7S,11S,Z)-11-hydroxy-7-(3′-(2″-methoxyethoxymethoxymethyl)-1′-methylpent-1′-enyl)-2,3-bis(methoxymethoxy)-12,12-dimethyl-6-oxabicyclo[11.3.1]heptadeca-1(17),13,15-trien-5,9-dione

Formula: C₃₃H₅₂O₁₁

Molar Mass: 624.76 g/mol

R_(f): 0.26 (pentane/acetone 7/3)

ESI-MS: 642.3 [M+NH₄ ⁺]

4.13 Synthesis of Analogue J51

To a cooled (0° C.) solution of J50 (5 mg, 8.0 μmol, 1.00 equivalent) in2.4 ml THF was added aqueous HCl (4M, 2.4 ml). After stirring 20 h atroom temperature, the mixture was diluted with 1 ml ethyl acetate,neutralized to pH 8 with sat. aq. NaHCO₃ and extracted with ethylacetate (5×15 ml). The combined organic extracts were dried over MgSO₄and concentrated in vacuo. The residue was purified HP-LC purification(LUNA C18, 0% to 100% acetonitrile in aqueous 5 nM ammonium acetate in30 min) provided J51 (0.6 mg, 17% over two steps) as a white solid.

(3R,3′R,4S,7S,11S,Z)-3,4,11-trihydroxy-7-(3′-hydroxymethyl-1′-methylpent-1′-enyl)-12,12-dimethyl-6-oxabicyclo[11.3.1]heptadeca-1(17),13,15-trien-5,9-dione

Formula: O₂₅H₃₆O₇

Molar Mass: 448.55 g/mol

ESI-MS: 449.2 [M+H⁺]

1H NMR (CHLOROFORM-d, 500 MHz): δ=7.33-7.39 (m, 1H), 7.28-7.32 (m, 1H),7.14 (br. d, J=7.3 Hz, 1H), 7.01-7.05 (m, 1H), 5.40 (dd, J=8.7, 3.4 Hz,1H), 5.01 (d, J=10.5 Hz, 1H), 4.17-4.25 (m, 2H), 3.62-3.66 (m, 1H),3.54-3.60 (m, 1H), 3.29 (app. t, J=10.1 Hz, 1H), 3.21 (dd, J=13.4, 4.0Hz, 1H), 2.88-2.93 (m, 2H), 2.75-2.82 (m, 1H), 2.71 (dd, J=18.3, 4.0 Hz,1H), 2.35-2.45 (m, 1H), 2.30 (dd, J=18.3, 8.2 Hz, 1H), 2.04-2.12 (m,1H), 1.67-1.76 (m (containing a doublet, 1.70), 4H), 1.57 (s, 3H), 1.47(s, 3H), 1.33-1.42 (m, 1H), 1.05-1.17 (m, 1H), 0.83 ppm (app. t, J=7.5Hz, 3H)

Example 5 Activity Tests of Phenyl Derivatives of PelorusideExperimental Procedures: Compounds

The test compound JC168 was dissolved in dimethyl sulfoxide (DMSO) at aconcentration of 0.001, 0.01, 0.1 or 1 μg/ml.

The microtubule inhibitor paclitaxel was used as a positive control. Itwas dissolved in physiologic saline solution (at a concentration of 0.1or 1 μg/ml).

Cell Lines MO4 Cells:

As test cells for functional assays MO4 cells were used, which aremurine virally transformed fibrosarcoma-like cells. This choice is basedon the following elements. These cells:

1. are highly directionally motile and invasive both in vitro and invivo2. are sensitive to microtubule inhibitors, and survive the M-block.3. are tumorigenic in C3H syngeneic mice.

MDA-MB231 GFP-LUC Cells:

Because MO4 cells do not invade when seeded on top of collagen, anothercell line was used for performing the collagen invasion assay, namelyMDA-MB232 GFP-LUC cells. These cells are derived from a human mammarycarcinoma and were stably transfected with green fluorescent protein(GFP) and luciferase (LUC).

PtK2 Cells

The PtK2 cell line was established from the kidney tissue of an adultmale rat kangaroo (Potorous tridactylus).

Functional Assays

The effect of the peloruside analog JC168 was tested on the followingfunctional assays in vitro:

1. Immunocytochemistry of the Cytoplasmic Microtubule Complex

Cells were seeded on glass coverslips in culture medium, supplementedwith fetal bovine serum, at an initial concentration of10,000/coverslip. After incubation overnight to allow spreading,treatments started at 37° C. for 1, 4 or 24 h. Cultures were then rinsedwith phosphate-buffered saline and fixed in methanol at −20° C. Theprimary rabbit anti-tubulin polyclonal antibody was a gift from Dr. DeBrabander (Janssen Life Sciences, Beerse, Belgium). Secondary antibodieswere goat anti-rabbit, and revealed either with a chromogen or afluorophor (Dako, Roskilde, Denmark).

2. Sulforhodamine B Assay

The sulforhodamine B (SRB) assay was used for cell densitydetermination. The assay is based on the measurement of cellular proteincontent (“biomass”). The method allows a large number of samples to betested within a few days, and is a quantitative assay for growthinhibition by microtubule inhibitors. The method has been optimized forthe screening of compounds for growth effects on adherent cells in a96-well format. After an incubation period, cell monolayers were fixedwith 10% (wt/vol) trichloroacetic acid and stained for 30 min, afterwhich the excess dye was removed by washing repeatedly with 1% (vol/vol)acetic acid. The protein-bound dye was dissolved in 10 mM Tris basesolution for OD determination at 510 nm using a microplate reader. Theresults were linear over a 20-fold range of cell numbers, and thesensitivity was comparable to those of fluorometric methods.

3. MTT Assay

MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, ayellow tetrazole), was reduced to purple formazan in living cells. Asolubilization solution (dimethyl sulfoxide) was added to dissolve theinsoluble purple formazan product into a purple solution. The absorbanceof this colored solution was quantified by measuring it at a wavelengthbetween 500 and 600 nm by a spectrophotometer. The absorption maximum isdependent on the solvent employed. The reductions take place only whenreductase enzymes are active, and therefore conversion is often used asa measure of viable (living) cells. However, it is important to keep inmind that other viability tests (such as the CASY cell countingtechnology) sometimes give different results, as many differentconditions apart from growth interference can increase or decreasemetabolic activity. As stated in another way, changes in metabolicactivity can give large changes in MTT results, while the number ofviable cells is constant. When the amount of purple formazan produced bycells treated with an agent is compared with the amount of formazanproduced by untreated control cells, the effectiveness of the agent incausing death or in changing the metabolism of cells can be deducedthrough the production of a dose-response curve.

4. Collagen Type I Invasion Assay

Tissue invasion requires infiltration into an extracellular matrix (ECM)dominated by networks of collagen type I. The invasion model, presentedhere, consists of native, acid-extracted rat tail collagen type Icontaining nonhelical telopeptides situated at the N- and C-terminalends. These telopeptides play an important role in intermolecularcovalent cross-links necessary for a gel architecture presenting itselfas a structural barrier to cancer cell traffic. Collagen type I solutionwas prepared with a final concentration of 1 mg ml⁻¹ collagen type I bymixing the following pre-cooled (stored at 4° C.) components: 4 volumescollagen type I (stock is 3.49 mg ml⁻¹), 5 volumes of calcium- andmagnesium-free Hank's balanced salt solution (CMF-HBSS), 1 volume ofminimal essential medium (MEM) (10×), 1 volume of 0.25 M NaHCO₃, 2.65volumes of standard medium and 0.3 volumes of 1M NaOH to make thesolution alkaline. The collagen type I solution was gently poured intothe wells of a 6-well plate. The experimental set-up was placed at 37°C. in a humidified atmosphere with 10% CO2 in air for at least 1 h.After gelification, a cell suspension of MDA-MB231 GFP-LUC cells wasadded on top of the collagen gels and incubated for 24 hours. Invasionof cells was observed in the transparent 3D collagen gels by phasecontrast microscopy as cells with extensions penetrating into thecollagen gel (FIG. 5B-D). Invasion is calculated as the percentage ofinvading cells per high powered field and is expressed as the mean andstandard deviation. The number of examined fields is 10. The sectionswere stained with hematoxylin-eosin.

5. Chick Heart Invasion Assay

Aggregates of MO4 cells were prepared by diluting single dissociatedcells to appropriate concentrations in 6 ml complete growth medium in a50 ml Erlenmeyer flask and incubated on a Gyrotory shaker at 37° C. and70 rpm in a humidified atmosphere with 10% CO2 in air for 72 h. Theaggregates were confronted overnight on top of semi-solid agar mediumwith precultured heart fragments (PHF, diameter 0.4 mm) prepared from9-day-old chick embryos. Suspension organotypic cultures were incubatedin 1.5 ml culture medium with or without a test compound on a Gyrotoryshaker at 120 rpm under a controlled atmosphere containing 10% CO2 inair. Cells were fixed in Bouin-Hollande's solution and embedded inparaffin for histologic determination of cell invasiveness. Consecutivesections were stained with haematoxylin-eosin. Invasion is defined asthe progressive occupation of PHF by the confronting test cells.Microscopic analysis of all consecutive sections from a confrontingculture allowed the reconstruction of the interaction between the cellaggregate and the PHF in three dimensions.

The observation of different patterns of interaction has led to thefollowing scale:

Grade 0: Only PHF is found. No confronting cells can be observed.Grade I: The confronting test cells are attached to the PHF, and do notoccupy the heart tissue, even not the outermost cell layers.Grade IIa: Occupation of the PHF is limited to the outer fibroblast-likeand myoblast cell layersGrade IIb: The PHF has surrounded the cell aggregate without signs ofoccupation.Grade III: The confronting cells have occupied the PHF, but have leftmore than half of the original amount of heart tissue intact.Grade IV: The confronting cells have occupied more than half of theoriginal volume of the PHF.

Grade I and II are observed with non-invasive cell populations, whilegrade III an IV are typical of invasion. To evaluate progression withtime, histological analysis was done on confronting cultures fixed afterdifferent incubation periods.

Results: 1. Immunocytochemistry of the Cytoplasmic Microtubule Complex

PtK2 cells were treated with DMSO (solvent control), JC168, orpaclitaxel (positive control). After 1 hour, 4 hours, and 24 hours ofincubation, cells were fixed and stained for the cytoplasmic microtubulecomplex.

TABLE 1 Effects of JC168 and paclitaxel on the cytoplasmic microtubulecomplex. Concentration Treatment (μg/ml) 1 hour 4 hours 24 hours DMSO —normal normal normal JC168 0.1 disturbed disturbed disturbed 1.0disturbed disturbed disturbed paclitaxel 0.1 normal disturbed disturbed1.0 disturbed disturbed disturbed JC168-treated PtK2 cells showed adisturbed microtubule complex similar to paclitaxel-treated cells. Theunordered cytoplasmic microtubule complex was already observed after 1hour of incubation with JC168.

2. Sulforhodamine B Assay

MO4 cells were treated with DMSO (solvent control), JC168, orpaclitaxel. After 4 days of incubation, a sulforhodamine B assay wasperformed to evaluate the effect of the treatments on cell growth.

FIG. 1 shows that JC168 inhibits cell growth. The potency of JC168 is 5%as compared to paclitaxel (FIG. 2).

3. MTT Assay

MO4 cells were treated with DMSO (solvent control), JC168, orpaclitaxel. After 4 days of incubation, a MTT assay was performed toevaluate the effect of the treatments on the viability of the cells, inparticular their metabolic activity, more in particular theirdehydrogenase activity.

FIG. 3 shows that JC168 inhibits the conversion of3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) intoformazan. This reduction in metabolic activity of jc163-treated MO4cells is mainly due to the growth-inhibiting effect of jc163 as revealedby the SRB assay (FIGS. 1-2).

The potency of jc163 to inhibit MTT conversion is 3% as compared topaclitaxel (FIG. 4).

4. Collagen Invasion Assay

MDA-MB231 GFP-LUC cells were treated with DMSO (solvent control), JC168,or paclitaxel (positive control) or left untreated (control) andincubated on a collagen type I gel. After 24 hours of incubation,invasion into the collagen was evaluated by phase-contrast microscopy.JC168 inhibits invasion of MDA-MB231 GFP-LUC cells into collagen type Ias shown in FIG. 5. The potency of JC168 to inhibit invasion in collagentype I is 89% as compared to paclitaxel. The histological sections ofthe collagen gels after 14 days show a low number of cells on top of thegel for the paclitaxel and JC168 treatments, whereas for the solventcontrol numerous cells are found on top of and in the collagen gel. Fromthis we can conclude that the number of invaded cells is clearly lowerin the treated cultures.

5. Chick Heart Invasion Assay

MO4 cells were confronted with precultured heart fragments and theorganotypic cultures were incubated in suspension in the presence ofDMSO (solvent control), JC168 or paclitaxel (positive control) for 2-4days.

TABLE 2 Effect of JC168 (1 μg/ml) and paclitaxel (1 μg/ml) on invasionof MO4 cells into embryonic chick heart. n = number of organotypiccultures classified according to the indicated scale. Incubation timetreatment Grade I Grade II Grade III Grade IV 2 days DMSO n = 5 JC168 n= 5 3 days DMSO n = 2 n = 3 JC168 n = 5 4 days DMSO n = 5 JC168 n = 3 n= 3 4 days DMSO n = 5 paclitaxel n = 5 JC168 inhibits invasion of MO4cells into embryonic chick heart tissue, but is less potent ininhibiting invasion as compared to paclitaxel (Table 2).

Example 6 Testing Pharmacological Efficacy

In order to evaluate the usefulness of the peloruside analog as amicrotubule inhibitor, following additional experiments are performedfor the compounds as defined herein:

(1) Test tube experiments to determine the binding affinity between thecompound and its molecular target.(2) Functional tests in vitro to assess the effects of the compound ongrowth, directional migration and invasion of malignant cellpopulations. Proposed assays are: (2a) Growth curve determinations viacell counts, sulforhodamine B biomass and MTT conversion measurements.(2b) Directional migration analysis via Boyden chamber and wound healingassays. (2c) Invasion assays in Matrigel, type I collagen gel andembryonic chick heart fragments.(3) Evaluation of the assays mentioned in (2) on different cell types:mammary, prostate, ovarian, colorectal, lung, melanoma and epidermoidcarcinoma as representatives of the most important human cancers, andfibroblasts, myofibroblasts and endothelial cells as representatives ofstromal cells.(4) Functional tests in laboratory nude mice to assess the effect ongrowth, invasion and metastasis formation. Proposed assays are: (4a)Assessment of growth, invasion, metastasis and angiogenesis offluorescently labeled tumor cells in the chick chorio-allantois membranetest. (4b) Volume measurements of heterotopically (subcutaneously)implanted tumor cells. (4c) Histology evaluation of invasion byorthotopically implanted cancer cells. (4d) Metastasis analysis ofluciferase-transfected tumor cells (injected intracardially) viabioluminescence detection.

1. A compound of Formula I:

wherein: X¹ is CR^(1a)R^(1b), X² is CR^(2a)R^(2b), X³ is CR^(3a)R^(3b),X⁴ is CR^(4a)R^(4b), and wherein: R^(1a) is selected from hydrogen,hydroxyl, halogen, and a group selected from —NR¹⁰R¹¹, C₁₋₆alkyl,C₂₋₆alkenyl, C₂₋₆alkynyl, C₆₋₁₀aryl, C₆₋₁₀arylC₁₋₆alkyl and C₁₋₆alkoxy,each group being independently optionally substituted with one or moresubstituent(s) each independently selected from hydroxyl, halogen,C₁₋₆alkyl and C₁₋₆alkoxy; R^(2a) is selected from hydrogen, hydroxyl,halogen, and a group selected from —NR¹⁰R¹¹, C₁₋₆alkyl, C₂₋₆alkenyl,C₂₋₆alkynyl, C₆₋₁₀aryl, C₆₋₁₀arylC₁₋₆alkyl and C₁₋₆alkoxy, each groupbeing independently optionally substituted with one or moresubstituent(s) each independently selected from hydroxyl, halogen,C₁₋₆alkyl and C₁₋₆alkoxy, R^(3a) is selected from hydrogen, hydroxyl,halogen, and a group selected from —NR¹⁰R¹¹, C₁₋₆alkyl, C₂₋₆alkenyl,C₂₋₆alkynyl, C₆₋₁₀aryl, C₆₋₁₀arylC₁₋₆alkyl and C₁₋₆alkoxy, each groupbeing independently optionally substituted with one or moresubstituent(s) each independently selected from hydroxyl, halogen,C₁₋₆alkyl and C₁₋₆alkoxy R^(4a) is selected from hydrogen, hydroxyl,halogen, and a group selected from —NR¹⁰R¹¹, C₁₋₆alkyl, C₂₋₆alkenyl,C₂₋₆alkynyl, C₆₋₁₀aryl, C₆₋₁₀arylC₁₋₆alkyl and C₁₋₆alkoxy, each groupbeing independently optionally substituted with one or moresubstituent(s) each independently selected from hydroxyl, halogen,C₁₋₆alkyl and C₁₋₆alkoxy; and wherein: R^(1b) is selected from hydrogen,hydroxyl, halogen, and a group selected from —NR¹⁰R¹¹, C₁₋₆alkyl,C₂₋₆alkenyl, C₂₋₆alkynyl, C₆₋₁₀aryl, C₆₋₁₀arylC₁₋₆alkyl and C₁₋₆alkoxy,each group being independently optionally substituted with one or moresubstituent(s) each independently selected from hydroxyl, halogen,C₁₋₆alkyl and C₁₋₆alkoxy, R^(2b) is selected from hydrogen, hydroxyl,halogen, and a group selected from —NR¹⁰R¹¹, C₁₋₆alkyl, C₂₋₆alkenyl,C₂₋₆alkynyl, C₆₋₁₀aryl, C₆₋₁₀arylC₁₋₆alkyl and C₁₋₆alkoxy, each groupbeing independently optionally substituted with one or moresubstituent(s) each independently selected from hydroxyl, halogen,C₁₋₆alkyl and C₁₋₆alkoxy, R^(3b) is selected from hydrogen, hydroxyl,halogen, and a group selected from —NR¹⁰R¹¹, C₁₋₆alkyl, C₂₋₆alkenyl,C₂₋₆alkynyl, C₆₋₁₀aryl, C₆₋₁₀arylC₁₋₆alkyl and C₁₋₆alkoxy, each groupbeing independently optionally substituted with one or moresubstituent(s) each independently selected from hydroxyl, halogen,C₁₋₆alkyl and C₁₋₆alkoxy, R^(4b) is selected from hydrogen, hydroxyl,halogen, and a group selected from —NR¹⁰R¹¹, C₁₋₆alkyl, C₂₋₆alkenyl,C₂₋₆alkynyl, C₆₋₁₀aryl, C₆₋₁₀arylC₁₋₆alkyl and C₁₋₆alkoxy, each groupindependently being independently optionally substituted with one ormore substituent(s) each independently selected from hydroxyl, halogen,C₁₋₆alkyl and C₁₋₆alkoxy; or wherein R^(1a) and R^(1b), or R^(2a) andR^(2b), or R^(3a) and R^(3b), or R^(4a) and R^(4b) taken togetherrepresent an oxo (═O) group; and wherein the bond represented by adashed and solid line represents a single bond or a double bond andwherein in case of a double bond, R^(1b) and R^(2b) are absent and atleast one of R^(1a) and R^(2a) is not OH or —NR¹⁰R¹¹; and wherein: R¹⁰and R¹¹ are each independently selected from hydrogen and C₁₋₆alkyl; andwherein: when R^(1a), R^(2a), R^(3a) or R^(4a) is hydroxyl, thecorresponding R^(1b), R^(2b), R^(3b) or R^(4b) is not hydroxyl, is not—NR¹⁰R¹¹, is not halogen, or is not C₁₋₆alkoxy, when R^(1a), R^(2a),R^(3a) or R^(4a) is —NR¹⁰R¹¹, the corresponding R^(1b), R^(2b), R^(3b)or R^(4b) is not hydroxyl, is not —NR¹⁰R¹¹, is not halogen, or is notC₁₋₆alkoxy, when R^(1a), R^(2a), R^(3a) or R^(4a) is halogen, thecorresponding R^(1b), R^(2b), R^(3b) or R^(4b) is not hydroxyl, or isnot —NR¹⁰R¹¹, when R^(1b), R^(2b), R^(3b) or R^(4b) is hydroxyl, thecorresponding R^(1a), R^(2a), R^(3a) or R^(4a) is not hydroxyl, is not—NR¹⁰R¹¹, is not halogen, or is not C₁₋₆alkoxy, and when R^(1b), R^(2b),R^(3b) or R^(4b) is —NR¹⁰R¹¹, the corresponding R^(1a), R^(2a), R^(3a)or R^(4a) is not hydroxyl, is not —NR¹⁰R¹¹, is not halogen, or is notC₁₋₆alkoxy. when R^(1b), R^(2b), R^(3b) or R^(4b) is halogen, thecorresponding R^(1a), R^(2a), R^(3a) or R^(4a) is not hydroxyl, or isnot —NR¹⁰R¹¹, and the stereoisomers, prodrugs, tautomers, racemates,salts, hydrates, or solvates thereof.
 2. The compound according to claim1, wherein R^(1b), R^(2b), R^(3b), and R^(4b) each independently areselected from hydrogen, halogen, and a group selected from C₁₋₆alkyl,C₂₋₆alkenyl, C₂₋₆alkynyl, C₆₋₁₀aryl, C₆₋₁₀arylC₁₋₆alkyl, and C₁₋₆alkoxy,each group being optionally substituted with one or more substituent(s)selected from hydroxyl, halogen, C₁₋₆alkyl, or C₁₋₆alkoxy; or whereinR^(1a) and R^(1b), and/or R^(2a) and R^(2b), and/or R^(3a) and R^(3b),and/or R^(4a) and R^(4b) taken together form an oxo (═O) group.
 3. Thecompound according to claim 1, having structural Formula Ia,

wherein R^(1b), R^(2b), R^(3b), and R^(4b) are each independentlyselected from hydrogen, hydroxyl, halogen, and a group selected from—NR¹⁰R¹¹, C₁₋₆alkyl, C₂₋₆alkenyl, C₂₋₆alkynyl, C₆₋₁₀aryl,C₆₋₁₀arylC₁₋₆alkyl and C₁₋₆alkoxy, each group independently beingoptionally substituted with one or more substituent(s) eachindependently selected from hydroxyl, halogen, C₁₋₆alkyl and C₁₋₆alkoxy;R^(1b) and R^(2b) each independently can represent an oxo (═O) group, inwhich case the bond represented by a dashed and solid line is a singlebond and the geminal hydrogen is absent; R^(3b) and R^(4b) eachindependently can represent an oxo (═O) group, in which case the geminalhydrogen is absent; and wherein when the bond represented by a dashedand solid line is a double bond with E- or Z-geometry, at least one ofR^(1b) and R^(2b) is not hydroxyl or —NR¹⁰R¹¹; and the stereoisomers,prodrugs, tautomers, racemates, salts, hydrates, or solvates thereof. 4.The compound according to claim 3, having structural Formula Ib, Ic orId,

wherein R^(1b), R^(2b), R^(3b), and R^(4b) have the same meaning asdefined in claim 2, and wherein in structure Ic, the double bond canhave the E- or Z-geometry.
 5. The compound according to claim 3, havingstructural Formula Ie, If, or Ig,

wherein R^(1b), R^(2b), R^(3b), and R^(4b) have the same meaning asdefined in claim 2, and the stereoisomers, prodrugs, tautomers,racemates, salts, hydrates, or solvates thereof.
 6. The compoundaccording to claim 3, having structural Formula Ih, Ii or Ij,

wherein R^(1b), R^(2b), R^(3b), and R^(4b) have the same meaning asdefined in claim 2, and the stereoisomers, prodrugs, tautomers,racemates, salts, hydrates, or solvates thereof.
 7. The compoundaccording to claim 1, wherein R^(3b) and R^(4b) have ananti-stereorelationship.
 8. The compound according to claim 1, whereinR^(1b) and R^(2b) are each independently selected from hydrogen,hydroxyl, —NR¹⁰R¹¹, halogen, C₁₋₆alkyl, and C₁₋₆alkoxy, preferablyR^(1b) and R^(2b) are hydrogen, halogen, hydroxyl or methoxy, or whereinR^(1b) is hydroxyl and R^(2b) is methoxy or vice versa, or whereinR^(1b) is halogen and R^(2b) is methoxy, or vice versa, or whereinR^(1b) is halogen and R^(2b) is hydroxyl, or vice versa, or whereinR^(1b) and R^(2b) are halogen, or wherein R^(1b) and R^(2b) are methoxy,or wherein R^(1b) and R^(2b) are hydroxyl.
 9. The compound according toclaim 1, wherein R^(3b) is selected from hydrogen, hydroxyl, —NR¹⁰R¹¹,halogen, C₁₋₆alkyl, and C₁₋₆alkoxy, or wherein R^(3b) is hydroxyl,methoxy or halogen.
 10. The compound according to claim 1, whereinR^(4b) is selected from the group consisting of hydrogen, hydroxyl,—NR¹⁰R¹¹, halogen, C₁₋₆alkyl, and C₁₋₆alkoxy, or wherein R^(4b) ishydroxyl, methoxy or halogen.
 11. The compound according to claim 1,having one of the following structural formulas:


12. A composition comprising a compound according to claim 1, and apharmaceutically acceptable excipient, and optionally comprising afurther active pharmaceutical ingredient.
 13. The compound according toclaim 1, for use as a medicament.
 14. The compound according to claim 1,for use in the treatment of a disease related to microtubule stability,or for use in the treatment of a proliferative disorder.
 15. A processfor producing a compound according to claim 1, comprising the steps of:(a) reacting a methyl ketone having structural Formula IIa with analdehyde having structural Formula IIb, through aldol coupling, whereinP⁴ and P⁵ are protecting groups, and executing suitable functional groupinterconversions, thereby obtaining a compound having structural FormulaII; (b) protecting the functional groups at positions X³ and X⁴ in thecompound of Formula II where needed; (c) reacting the compound ofFormula II with a compound of the Formula III in the presence of asuitable catalyst, giving rise to a compound of the formula (IV), (d)removing the protecting groups P¹ and P⁴ in the resulting compound andesterifying the deprotected COOH with the C₁₅ OH group, executingsuitable functional group interconversions, thereby obtaining thelactone having structural Formula V; and (e) deprotecting P⁵ in thecompound having structural Formula V and deprotecting any of thepossibly protected X¹, X², X³, or X⁴ groups, if required after executingadditional suitable functional group interconversions, thereby obtaininga compound having structural Formula I:

wherein P¹ is hydrogen or a carboxyl protecting group, preferably anoptionally substituted alkyl, more preferably methyl; P² is selectedfrom halogen, pseudohalogen, CF₃SO₃ and OAc, preferably P² is bromo,and, simultaneously, P³ is a trialkyltin, preferably trimethyltin ortri-n-butyltin, or P³ is boronic acid or a boronic ester; or,alternatively, wherein P² is a trialkyltin, preferably trimethyltin ortri-n-butyltin, or P³ is boronic acid or a boronic ester, andsimultaneously, P³ is selected from halogen, pseudohalogen, CF₃SO₃ andOAc, preferably P³ is bromo; P⁴ is selected from an orthogonally chosenprotecting group, preferably 4-OMe-Bn; P⁵ is a protecting group whichcan be orthogonally removed, preferably selected from: TBS and MEM; thecatalyst is a transition metal with ligands, preferably Pd₂(dba)₃.CHCl₃;and X¹, X², X³, and X⁴ have the same meaning as that defined in claim 1,and wherein, if present, their functional groups are suitably protectedin steps (a) and (b), functionally interconverted where needed anddeprotected accordingly in step (e).